WO2002059255A2

WO2002059255A2 - Protein-protein interactions in $i(saccharomyces cerevisiae)

Info

Publication number: WO2002059255A2
Application number: PCT/EP2002/001350
Authority: WO
Inventors: Pierre Legrain
Original assignee: Hybrigenics
Priority date: 2001-01-26
Filing date: 2002-01-25
Publication date: 2002-08-01

Abstract

The present invention relates to proteins that interact with other proteins of Saccharomyces cerevisiae. More specifically, the present invention relates to complexes of polypeptides or polynucleotides encoding the polypeptides, fragments of the polypeptides, antibodies to the complexes, Selected Interacting Domains (SID®) which are identified due to the protein-protein interactions, methods for screening for agents which modulate the interaction of proteins and compositions that are capable of modulating the protein-protein interactions, such as, for example, drug in pharmaceutical composition.

Description

PROTEIN-PROTEIN INTERACTIONS IN Saccharomyces cerevisiae

FIELD OF THE INVENTION

In another embodiment the present invention provides a protein-protein interaction map called a PIM® which is available in a report relating to the protein-protein interactions of Saccharomyces cerevisiae.

In yet another embodiment the present invention relates to the identification of additional proteins in the pathway common to the proteins described therein, such as proteasome, signal transduction, protein synthesis, apoptosis, cell cycle secretion and metabolic pathways. S. cerevisiae is also a research model organism for the study of Candida sp.

BACKGROUND AND PRIOR ART

Most biological processes involve specific protein-protein interactions. Protein- protein interactions enable two or more proteins to associate. A large number of non_¬ covalent bonds form between the proteins when two protein surfaces are precisely matched. These bonds account for the specificity of recognition. Thus, protein-protein interactions are involved, for example, in the assembly of enzyme subunits, in antibody- antigen recognition, in the formation of biochemical complexes, in the correct folding of proteins, in the metabolism of proteins, in the transport of proteins, in the localization of proteins, in protein turnover, in first translation modifications, in the core structures of viruses and in signal transduction.

General methodologies to identify interacting proteins or to study these interactions have been developed. Among these methods are the two-hybrid systems originally developed by Fields and co-workers and described, for example, in U.S. Patent Nos. 5,283,173, 5,468,614 and 5,667,973, which are hereby incorporated by reference.

The earliest and simplest two-hybrid system, which acted as basis for development of other versions, is an in vivo assay between two specifically constructed proteins. The first protein, known in the art as the "bait protein" is a chimeric protein which binds to a site on DNA upstream of a reporter gene by means of a DNA-binding domain or BD. Commonly, the binding domain is the DNA-binding domain from either Gal4 or native E. coli LexA and the sites placed upstream of the reporter are Gal4 binding sites or LexA operators, respectively.

The second protein is also a chimeric protein known as the "prey" in the art. This second chimeric protein carries an activation domain or AD. This activation domain is typically derived from Gal4, from VP16 or from B42.

Besides the two hybrid systems, other improved systems have been developed to detected protein-protein interactions. For example, a two-hybrid plus one system was developed that allows the use of two proteins as bait to screen available cDNA libraries to detect a third partner. This method permits the detection between proteins that are part of a larger protein complex such as the RNA polymerase ll holoenzyme and the TFIIH or TFIID complexes. Therefore, this method, in general, permits the detection of ternary complex formation as well as inhibitors preventing the interaction between the two previously defined fused proteins. Another advantage of the two-hybrid plus one system is that it allows or prevents the formation of the transcriptional activator since the third partner can be expressed from a conditional promoter such as the methionine-repressed Met25 promoter which is positively regulated in medium lacking methionine. The presence of the methionine-regulated promoter provides an excellent control to evaluate the activation or inhibition properties of the third partner due to its "on" and "off¹ switch for the formation of the transcriptional activator. The three-hybrid method is described, for example in Tirode et al., The Journal of Biological Chemistry, 272, No. 37 pp. 22995-22999 (1997). incorporated herein by reference.

Besides the two and two-hybrid plus one systems, yet another variant is that described in Vidal et al, Proc. Natl. Sci. 93 pgs. 10315-10320 called the reverse two- and one-hybrid systems where a collection of molecules can be screened that inhibit a specific protein-protein or protein/DNA interactions, respectively.

A summary of the available methodologies for detecting protein-protein interactions is described in Vidal and Legrain, Nucleic Acids Research Vol. 27, No. 4 pgs.919-929 (1999) anc Legrain and Selig, FEBS Letters 480 pgs. 32-36 (2000) which references are incorporated herein by reference.

However, the above conventionally used approaches and especially the commonly used two-hybrid methods have their drawbacks. For example, it is known in the art that, more often than not, false positives and false negatives exist in the screening method. In fact, a doctrine has been developed in this field for interpreting the results and in common practice an additional technique such as co-immunoprecipitation or gradient sedimentation of the putative interactors from the appropriate cell or tissue type is generally performed. The methods used for interpreting the results are described by Brent and Finley, Jr. in Ann. Rev. Genet, 31 pgs. 663-704 (1997). Thus, the data interpretation is very questionable using the conventional systems.

One method to overcome the difficulties encountered with the methods in the prior art is described in WO 99/42612, incorporated herein by reference. This method is similar to the two-hybrid system described in the prior art in that it also uses bait and prey polypeptides. However, the difference with this method is that a step of mating at least one first haploid recombinant yeast cell containing the prey polypeptide to be assayed with a second haploid recombinant yeast cell containing the bait polynucleotide is performed. Of course the person skilled in the art would appreciate that either the first recombinant yeast cell or the second recombinant yeast cell also contains at least one detectable reporter gene that is activated by a polypeptide including a transcriptional activation domain.

The method described in WO 99/42612 permits the screening of more prey polynucleotides with a given bait polynucleotide in a single step than in the prior art systems due to the cell to cell mating strategy between haploid yeast cells. Furthermore, this method is more thorough and reproducible, as well as sensitive. Thus, the presence of false negatives and/or false positives is extremely minimal as compared to the conventional prior art methods.

Yeast are unicellular eukaryotic organisms; the yeast genus Saccharomyces comprises strains whose biochemistry and genetics are intensively studied in the laboratory; it also comprises strains frequently used in the industry, in particular in the food industry (bread, alcoholic drinks, etc.), and consequently produced in very large quantities.

Among the numerous applications involving Saccharomyces cerevisiae, the most frequent are:

(1 ) Bread production.

(2) Production of ethanol or ethyl alcohol (C2-H5-OH) by fermentation methods.

Fermentation techniques for ethanol production developed during the early part of this century were supplemented by synthetic processes based on crude petroleum, as oil was much cheaper and abundantly available. However, the fermentative production of ethanol has again picked up, using various kinds of renewable fermentable substrates, such as: (i) sugar (from sugar-cane, sugar beet, fruit) which may be converted to ethanol directly; (ii) starch (from grain, root crops) which is first hydrolyzed to fermentable sugars by enzymes; and (iii) cellulose (from wood, agricultural wastes, etc.) which is converted to sugars. (Biotechnology: Economic and Social Aspects-Issues for Developing Countries, Eds. E. J. Da Silva, C. Ratiedge and A Sesson; Cambridge University Press, p.24, 1992).

Ethanol production by fermentation is based mainly on yeast, and for large scale fuel production, these are generally of the genus Saccharomyces.

(3) Additives

Yeast extract may provide product containing a large quantity of enzymes, coenzymes, ferments, group B vitamins, nucleotides, nucleosides, free amino acids and RNA acid. Particularly useful products are obtained from Saccharomyces cerevisiae strains having high resistance in an acid environment (e g. gastric juices) and towards antibiotics. These product characteristics make it particularly suitable and effective as a human and animal food additive, as a growth factor and intestinal bacterial flora regulator. Its action is both prophylactic and curative in many affections in the human and veterinary field deriving from enzymatic and bacterial imbalance of the intestine.

(4) Production of proteins

The ease with which the genetics of Saccharomyces cerevisiae cells may be manipulated and the long industrial history of this species hence make it a host of choice for the production of foreign proteins using recombinant DNA techniques.

Yeast cells have proven useful as hosts for production of heterologous gene products. Yeast such as the bakers yeast Saccharomyces cerevisiae can be grown to high cell densities inexpensively in simple media, and helpful genetic techniques and molecular genetic methods are available. Accordingly, pharmaceutical preparations of human alpha-1 -antitrypsin, and vaccines for hepatitis B virus have been produced in the cytoplasm of yeast cells and isolated by lysis of cells and purification of the desired protein (Valenzueia, P., et al., 1982, Nature 298: 347-350; Travis, J., et al, 1985, J. Biol. Chem. 260: 4384-4389). However, some proteins, such as prochymosin and prourokinase (also known as single-chain urinary plasminogen activator, or scu-PA) are produced much more efficiently by secretion from yeast cells, apparently because they are normally secreted from their native host cells and because proper folding of the polypeptide chain and disulfide bond formation occur only in the secretion pathway (Smith, Duncan, & Moir, 1985, Science 229: 1219-1224; Moir et al., 1988, Abstract 19 from The Ninth International Congress on Fibrinolysis, Amsterdam, The Netherlands).

This application needs improvement especially concerning the secretion of the protein of interest: The secreted yield of protein product is dependent upon both the gene to be expressed and the promoter and signal sequences chosen for its expression (Hitzeman, R. A., Leung, D. W., Perry, L. J., Kohr, W. J., Levine, H. L. and Goeddel, D. V. (1983) Science 219, 620-625; Bitter, G. A., Chen, K. K., Banks, A. R. and Lai, P.-H. (1984) Proc. Natl. Acad. Sci. USA 81 , 5330-5334; Brake, A. J., Merryweather, J. P., Coit, D. G., Heberlein, U. A., Masiarz, F. R., Mullenbach, G. T., Urdea, M. S., Valenzuela, P. and Barr, P. J. (1984) Proc. Natl. Acad. Sci. USA 81 , 4642-4646; Brake, A. J., Cousens, L. S., Urdea, M. S., Valenzuela, P. D. T. (1984) European Patent Application Publication No. 0 121 884). Although it is usually possible to obtain reasonably good production levels for a particular protein, often only a small fraction of the total amount produced can actually be found free in the medium. Most of the protein remains trapped inside the cell, often in the intracellular vacuole found in this species. In yeast, secretion can be regarded as a branched pathway with some secreted yeast proteins being "secreted" into the vacuole and others being directed across the plasma membrane to the periplasm and beyond (Sheckman, R. and Novick, P., in Strathem, J. N., Jones, E. W. and Broach, J. R. (eds.), Molecular Biology of the yeast Saccharomyces cerevisiae, Cold Springs Harbor Laboratory, Cold Springs Harbor, New York, 1981 , pp. 361-398). Apparently, some protein products of foreign genes are directed into the vacuolar branch of this pathway.

Yields of secreted heterologous proteins from yeast fermentations have been limited. Most non-yeast proteins are secreted quite inefficiently from yeast cells. For example, in all of the following cases, at least as much of the heteroloqous protein is found inside the cell as is found outside the cell in the culture broth or between the cell membrane and wall. This is true for calf prochymosin (Smith, Duncan, & Moir, 1985, Science 229: 1219-1223), human alpha-1-antitrypsin (Moir & Dumais, 1987, Gene 56: 209-217), human tissue plasminogen activator (Lemontt et al., 1985, DNA 4: 419-428), anchor-minus influenza hemagglutinin (Jabbar & Nayak, 1987, Mol. Cell. Biol. 7: 1476- 1485), alpha interferon (Hitzeman et al., 1983, Science 219: 620-625), a consensus interferon (Zsebo et al., 1986, J. Biol. Chem. 261 : 5858-5865), murine lambda and mu immunoglobulin chains (Wood et al., 1985, Nature 314: 446-449), and human lysozyme (Jigami et al., 1986, Gene 43: 273-279). Clearly, methods are needed to increase the efficiency of secretion of these proteins and other non-yeast proteins from yeast cells. Such methods would provide therapeutic and industrially useful proteins more economically.

(5) Model organism for medical study

S. cerevisiae is a research model organism for Candida infection study. It is also a model organism for human diseases, especially mechanisms involved in cancer (DNA repair, apoptose), neurodegenerative disease.

The two last fields of application request a precise knowledge of yeast pathways, that is why there is a great need for construction of protein interaction map (PIM®) of Saccharomyces cerevisiae and to identify protein function in pathways of interest.

The present invention relates to identifying protein-protein interactions for Saccharomyces cerevisiae.

The present invention also relates to identifying protein-protein interactions of Saccharomyces cerevisiae for the development of more effective and better targeted therapeutic applications, for the development of yeast strains having a better secretion yield of the protein of interest to produce (i. e., expression and secretion).

The present invention is also aimed at identifying complexes of polypeptides or polynucleotides encoding the polypeptides and fragments of the polypeptides of Saccharomyces cerevisiae. Also, the present invention relates to identifying antibodies to these complexes of polypeptides or polynucleotides encoding the polypeptides and fragments of the polypeptides of Saccharomyces cerevisiae including polyclonal, as well as monoclonal antibodies that are used for detection.

The present invention also concerns the identification of selected interacting domains of the polypeptides, called SID® polypeptides.

The present invention also relates to identifying selected interacting domains of the polynucleotides, called SID® polynucleotides.

Furthermore, the present invention is aimed at generating protein-protein interactions maps called PIM®s.

Also, the present invention concerns a method for screening drugs for agents which modulate the interaction of proteins and pharmaceutical compositions that are capable of modulating the protein-protein interactions of Saccharomyces cerevisiae.

The present invention also relates to administering the nucleic acids described herein via gene therapy.

Also, the present invention provides protein chips or protein microarrays.

In another embodiment, the present invention provides a report in, for example paper, electronic and/or digital forms, concerning the protein-protein interactions, the modulating compounds and the like as well as a PIM®.

SUMMARY OF THE PRESENT INVENTION

Thus the present invention relates to a protein complex of Saccharomyces cerevisiae. Furthermore, the present invention provides SID® polynucleotides and SID® polypeptides, as well as a PIM® for Saccharomyces cerevisiae.

The present invention also provides antibodies to the protein-protein complexes for Saccharomyces cerevisiae.

In another embodiment the present invention provides a method for screening drugs for agents that modulate the protein-protein interactions and pharmaceutical compositions that are capable of modulating protein-protein interactions.

In another embodiment the present invention provides protein chips or protein microarrays.

In yet another embodiment the present invention provides a report in, for example, paper, electronic and/or digital forms.

BRIEF DESCRIPTION OF THE DRAWINGS

Fig. 1 is a schematic representation of the pB1 plasmid. Fig. 2 s a schematic representation of the pB5 plasmid. Fig. 3 s a schematic representation of the pB6 plasmid. Fig. 4 s a schematic representation of the pB13 plasmid. Fig. 5 s a schematic representation of the pB14 plasmid. Fig. 6 is a schematic representation of the pB20 plasmid. Fig. 7 Is a schematic representation of the pP1 plasmid. Fig. 8 is a schematic representation of the pP2 plasmid. Fig. 9 s a schematic representation of the pP3 plasmid. Fig. 10 is a schematic representation of the pP6 plasmid. Fig. 1 1 is a schematic representation of the pP7 plasmid.

Fig. 12 is a schematic representation of vectors expressing the T25 fragment. Fig. 13 is a schematic representation of vectors expressing the T18 fragment. Fig. 14 is a schematic representation of various vectors of pCmAHLI , pT25 and pT18. Fig. 15 is a schematic representation identifying a SID®'s. In this figure the "Full- length prey protein" is the Open Reading Frame (ORF) or coding sequence (CDS) where the identified prey polypeptides are included. The Selected Interaction Domain (SID®) is determined by the commonly shared polypeptide domain of every selected prey fragment.

Fig. 16 is an example of protein interaction map (PIM®).

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

As used herein the terms "polynucleotides", "nucleic acids" and "oligonucleotides" are used interchangeably and include, but are not limited to RNA, DNA, RNA/DNA sequences of more than one nucleotide in either single chain or duplex form. The polynucleotide sequences of the present invention may be prepared from any known method including, but not limited to, any synthetic method, any recombinant method, any ex vivo generation method and the like, as well as combinations thereof.

The term "polypeptide" means herein a polymer of amino acids having no specific length. Thus, peptides, oligopeptides and proteins are included in the definition of "polypeptide" and these terms are used interchangeably throughout the specification, as well as in the claims. The term "polypeptide" does not exclude post-translational modifications such as polypeptides having covalent attachment of glycosyl groups, acetyl groups, phosphate groups, lipid groups and the like. Also encompassed by this definition of "polypeptide" are homologs thereof.

By the term "homologs" is meant structurally similar genes contained within a given species, orthologs are functionally equivalent genes from a given species or strain, as determined for example, in a standard complementation assay. Thus, a polypeptide of interest can be used not only as a model for identifying similar genes in given strains, but also to identify homologs and orthologs of the polypeptide of interest in other species. The orthologs, for example, can also be identified in a conventional complementation assay. In addition or alternatively, such orthologs can be expected to exist in bacteria (or other kind of cells) in the same branch of the phylogenic tree, as set forth, for example, at ftp : -' 'ftp . cme.msu. edu/pub/rdp/S S U-rRN A S S U/Prok.phylo . As used herein the term "prey polynucleotide" means a chimeric polynucleotide encoding a polypeptide comprising (i) a specific domain; and (ii) a polypeptide that is to be tested for interaction with a bait polypeptide. The specific domain is preferably a transcriptional activating domain.

As used herein, a "bait polynucleotide" is a chimeric polynucleotide encoding a chimeric polypeptide comprising (i) a complementary domain; and (ii) a polypeptide that is to be tested for interaction with at least one prey polypeptide. The complementary domain is preferably a DNA-binding domain that recognizes a binding site that is further detected and is contained in the host organism.

As used herein "complementary domain" is meant a functional constitution of the activity when bait and prey are interacting; for example, enzymatic activity.

As used herein "specific domain" is meant a functional interacting activation domain that may work through different mechanisms by interacting directly or indirectly through intermediary proteins with RNA polymerase II or Ill-associated proteins in the vicinity of the transcription start site.

As used herein the term "complementary" means that, for example, each base of a first polynucleotide is paired with the complementary base of a second polynucleotide whose orientation is reversed. The complementary bases are A and T (or A and U) or C and G.

The term "sequence identity" refers to the identity between two peptides or between two nucleic acids. Identity between sequences can be determined by comparing a position in each of the sequences which may be aligned for the purposes of comparison. When a position in the compared sequences is occupied by the same base or amino acid, then the sequences are identical at that position. A degree of sequence identity between nucleic acid sequences is a function of the number of identical nucleotides at positions shared by these sequences. A degree of identity between amino acid sequences is a function of the number of identical amino acid sequences that are shared between these sequences. Since two polypeptides may each (i) comprise a sequence (i.e., a portion of a complete polynucleotide sequence) that is similar between two polynucleotides, and (ii) may further comprise a sequence that is divergent between two polynucleotides, sequence identity comparisons between two or more polynucleotides over a "comparison window" refers to the conceptual segment of at least 20 contiguous nucleotide positions wherein a polynucleotide sequence may be compared to a reference nucleotide sequence of at least 20 contiguous nucleotides and wherein the portion of the polynucleotide sequence in the comparison window may comprise additions or deletions (i.e., gaps) of 20 percent or less compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two sequences.

To determine the percent identity of two amino acids sequences or two nucleic acid sequences, the sequences are aligned for optimal comparison. For example, gaps can be introduced in the sequence of a first amino acid sequence or a first nucleic acid sequence for optimal alignment with the second amino acid sequence or second nucleic acid sequence. The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, the molecules are identical at that position.

The percent identity between the two sequences is a function of the number of identical positions shared by the sequences. Hence % identity = number of identical positions / total number of overlapping positions X 100.

In this comparison the sequences can be the same length or may be different in length. Optimal alignment of sequences for determining a comparison window may be conducted by the local homology algorithm of Smith and Waterman (J. Theor. Biol., 91 (2) pgs. 370-380 (1981 ), by the homology alignment algorithm of Needleman and Wunsch, J. Miol. Biol., 48(3) pgs. 443-453 (1972), by the search for similarity via the method of Pearson and Lipman, PNAS, USA, 85(5) pgs. 2444-2448 (1988) , by computerized implementations of these algorithms (GAP, BESTFIT, FASTA and TFASTA in the Wisconsin Genetics Software Package Release 7.0, Genetic Computer Group, 575, Science Drive, Madison, Wisconsin) or by inspection. The best alignment (i.e., resulting in the highest percentage of identity over the comparison window) generated by the various methods is selected.

The term "sequence identity" means that two polynucleotide sequences are identical

(i.e., on a nucleotide by nucleotide basis) over the window of comparison. The term "percentage of sequence identity" is calculated by comparing two optimally aligned sequences over the window of comparison, determining the number of positions at which the identical nucleic acid base (e.g., A, T, C, G, U, or I) occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison (i.e., the window size) and multiplying the result by 100 to yield the percentage of sequence identity. The same process can be applied to polypeptide sequences.

The percentage of sequence identity of a nucleic acid sequence or an amino acid sequence can also be calculated using BLAST software (Version 2.06 of September 1998) with the default or user defined parameter.

The term "sequence similarity" means that amino acids can be modified while retaining the same function. It is known that amino acids are classified according to the nature of their side groups and some amino acids such as the basic amino acids can be interchanged for one another while their basic function is maintained.

The term "isolated" as used herein means that a biological material such as a nucleic acid or protein has been removed from its original environment in which it is naturally present. For example, a polynucleotide present in a plant, mammal or animal is present in its natural state and is not considered to be isolated. The same polynucleotide separated from the adjacent nucleic acid sequences in which it is naturally inserted in the genome of the plant or animal is considered as being "isolated."

The term "isolated" is not meant to exclude artificial or synthetic mixtures with other compounds, or the presence of impurities which do not interfere with the biological activity and which may be present, for example, due to incomplete purification, addition of stabilizers or mixtures with pharmaceutically acceptable excipients and the like.

"Isolated polypeptide" or "isolated protein" as used herein means a polypeptide or protein which is substantially free of those compounds that are normally associated with the polypeptide or protein in a naturally state such as other proteins or polypeptides, nucleic acids, carbohydrates, lipids and the like.

The term "purified" as used herein means at least one order of magnitude of purification is achieved, preferably two or three orders of magnitude, most preferably four or five orders of magnitude of purification of the starting material or of the natural material.

Thus, the term "purified" as utilized herein does not mean that the material is 100% purified and thus excludes any other material.

The term "variants" when referring to, for example, polynucleotides encoding a polypeptide variant of a given reference polypeptide are polynucleotides that differ from the reference polypeptide but generally maintain their functional characteristics of the reference polypeptide. A variant of a polynucleotide may be a naturally occurring allelic variant or it may be a variant that is known naturally not to occur. Such non-naturally occurring variants of the reference polynucleotide can be made by, for example, mutagenesis techniques, including those mutagenesis techniques that are applied to polynucleotides, cells or organisms.

Generally, differences are limited so that the nucleotide sequences of the reference and variant are closely similar overall and, in many regions identical.

Variants of polynucleotides according to the present invention include, but are not limited to, nucleotide sequences which are at least 95% identical after alignment to the reference polynucleotide encoding the reference polypeptide. These variants can also have 96%, 97%, 98% and 99.999% sequence identity to the reference polynucleotide. Nucleotide changes present in a variant polynucleotide may be silent, which means that these changes do not alter the amino acid sequences encoded by the reference polynucleotide.

Substitutions, additions and/or deletions can involve one or more nucleic acids.

Alterations can produce conservative or non-conservative amino acid substitutions, deletions and/or additions.

Variants of a prey or a SID® polypeptide encoded by a variant polynucleotide can possess a higher affinity of binding and/or a higher specificity of binding to its protein or polypeptide counterpart, against which it has been initially selected. In another context, variants can also loose their ability to bind to their protein or polypeptide counterpart.

By "anabolic pathway" is meant a reaction or series of reactions in a metabolic pathway that synthesize complex molecules from simpler ones, usually requiring the input of energy. An anabolic pathway is the opposite of a catabolic pathway.

As used herein, a "catabolic pathway" is a series of reactions in a metabolic pathway that break down complex compounds into simpler ones, usually releasing energy in the process. A catabolic pathway is the opposite of an anabolic pathway.

As used herein, "drug metabolism" is meant the study of how drugs are processed and broken down by the body. Drug metabolism can involve the study of enzymes that break down drugs, the study of how different drugs interact within the body and how diet and other ingested compounds affect the way the body processes drugs.

As used herein, "metabolism" means the sum of all of the enzyme-catalyzed reactions in living cells that transform organic molecules.

By "secondary metabolism" is meant pathways producing specialized metabolic products that are not found in every cell. As used herein, "SID®" means a Selected Interacting Domain and is identified as follows: for each bait polypeptide screened, selected prey polypeptides are compared. Overlapping fragments in the same ORF or CDS define the selected interacting domain.

As used herein the term "PIM®" means a protein-protein interaction map. This map is obtained from data acquired from a number of separate screens using different bait polypeptides and is designed to map out all of the interactions between the polypeptides.

The term "affinity of binding", as used herein, can be defined as the affinity constant Ka when a given SID® polypeptide of the present invention which binds to a polypeptide and is the following mathematical relationship: [SID®/polypeptide complex]

Ka =

[free SID®] [free polypeptide]

wherein [free SID®], [free polypeptide] and [SID®/polypeptide complex] consist of the concentrations at equilibrium respectively of the free SID® polypeptide, of the free polypeptide onto which the SID® polypeptide binds and of the complex formed between

SID® polypeptide and the polypeptide onto which said SID® polypeptide specifically binds.

The affinity of a SID® polypeptide of the present invention or a variant thereof for its polypeptide counterpart can be assessed, for example, on a Biacore™ apparatus marketed by Amersham Pharmacia Biotech Company such as described by Szabo et al Curr Opin Struct Bio\ 5 pgs. 699-705 (1995) and by Edwards and Leartherbarrow, Anal.

Biochem 246 pgs. 1-6 (1997).

As used herein the phrase "at least the same affinity" with respect to the binding affinity between a SID® polypeptide of the present invention to another polypeptide means that the Ka is identical or can be at least two-fold, at least three-fold or at least five fold greater than the Ka value of reference. As used herein, the term "modulating compound" means a compound that inhibits or stimulates or can act on another protein which can inhibit or stimulate the protein-protein interaction of a complex of two polypeptides or the protein-protein interaction of two polypeptides.

More specifically, the present invention comprises complexes of polypeptides or polynucleotides encoding the polypeptides composed of a bait polypeptide, or a bait polynucleotide encoding a bait polypeptide and a prey polypeptide or a prey polynucleotide encoding a prey polypeptide. The prey polypeptide or prey polynucleotide encoding the prey polypeptide is capable of interacting with a bait polypeptide of interest in various hybrid systems.

As described in the Background of the present invention there are various methods known in the art to identify prey polypeptides that interact with bait polypeptides of interest. These methods, include, but are not limited to, generic two-hybrid systems as described by Fields et al in Nature, 340:245-246 (1989) and more specifically in U.S. Patent Nos. 5,283,173, 5,468,614 and 5,667,973, which are hereby incorporated by reference; the reverse two-hybrid system described by Vidal et al, supra; the two plus one hybrid method described, for example, in Tirode et al, supra; the yeast forward and reverse 'n'-hybrid systems as described in Vidal and Legrain, supra; the method described in WO 99/42612; those methods described in Legrain et al FEBS Letters 480 pgs. 32-36 (2000) and the like.

The present invention is not limited to the type of method utilized to detect protein- protein interactions and therefore any method known in the art and variants thereof can be used. It is however better to use the method described in WO 99/42612 or WO 00/66722, both references incorporated herein by reference due to the methods' sensitivity, reproducibility and reliability.

Protein-protein interactions can also be detected using complementation assays such as those described by Pelletier et al. at http://\v\v\Λ-.abrf.ora/JBT Articles/JBTQ012/ibtOQ12.html. WO 00/07038 and WO98/34120. Although the above methods are described for applications in the yeast system, the present invention is not limited to detecting protein-protein interactions using yeast, but also includes similar methods that can be used in detecting protein-protein interactions in, for example, mammalian systems as described, for example in Takacs et al., Proc. Natl. Acad. Sci., USA, 90 (21 ):10375-79 (1993) and Vasavada et al., Proc. Natl. Acad. Sci., USA, 88 (23): 10686-90 (1991), as well as a bacterial two-hybrid system as described in Karimova et al (1998), WO99/28746, WO 00/66722 and Legrain et al FEBS Letters, 480 pgs. 32-36 (2000).

The above-described methods are limited to the use of yeast, mammalian cells and

Escherichia coli cells, the present invention is not limited in this manner. Consequently, mammalian and typically human cells, as well as bacterial, yeast, fungus, insect, nematode and plant cells are encompassed by the present invention and may be transfected by the nucleic acid or recombinant vector as defined herein.

Examples of suitable cells include, but are not limited to, VERO cells, HELA cells such as ATCC No. CCL2, CHO cell lines such as ATCC No. CCL61 , COS cells such as

COS-7 cells and ATCC No. CRL 1650 cells, W138, BHK, HepG2, 3T3 such as ATCC No.

CRL6361 , A549, PC12, K562 cells, 293 cells, Sf9 cells such as ATCC No. CRL1711 and Cv1 cells such as ATCC No. CCL70.

Other suitable cells that can be used in the present invention include, but are not limited to, prokaryotic host cells strains such as Escherichia coli, (e.g., strain DH5-α), Bacillus subtilis, Salmonella typhimurium, or strains of the genera of Pseudomonas, Streptomyces and Staphylococcus.

Further suitable cells that can be used in the present invention include yeast cells such as those of Saccharomyces such as Saccharomyces cerevisiae.

The bait polynucleotide, as well as the prey polynucleotide can be prepared according to the methods known in the art such as those described above in the publications and patents reciting the known method per se. The bait polynucleotide of the present invention is obtained from Saccharomyces cerevisiae genomic DNA, usually Open Reading Frame. The prey polynucleotide is obtained from Saccharomyces cerevisiae genomic DNA, variants of genomic DNA, and fragments from the genome or transcriptome of Saccharomyces cerevisiae ranging from about 200 nucleic acids to about 3000 nucleic acids. The prey polynucleotide is then selected, sequenced and identified.

A genomic DNA prey library is prepared from the Saccharomyces cerevisiae and constructed in the specially designed prey vector pP2 as shown in Figure 8 after ligation of suitable linkers such that every genomic DNA fragment insert is fused to a nucleotide sequence in the vector that encodes the transcription activation domain of a reporter gene. Any transcription activation domain can be used in the present invention. Examples include, but are not limited to, Gal4,YP16, B42, His and the like. Toxic reporter genes, such as CAT^R, CYH2, CYH1 , URA3, bacterial and fungi toxins and the like can be used in reverse two-hybrid systems.

The polypeptides encoded by the nucleotide inserts of the genomic DNA fragment prey library thus prepared are termed "prey polypeptides" in the context of the presently described selection method of the prey polynucleotides.

The bait polynucleotide can be inserted in bait plasmid pB6 as illustrated in Figure 3. The bait polynucleotide insert is fused to a polynucleotide encoding the binding domain of, for example, the Gal4 DNA binding domain and the shuttle expression vector is used to transform cells.

As stated above, any cells can be utilized in transforming the bait and prey polynucleotides of the present invention including mammalian cells, bacterial cells, yeast cells, insect cells and the like.

In an embodiment, the present invention identifies protein-protein interactions in yeast. In using known methods a prey positive clone is identified containing a vector which comprises a nucleic acid insert encoding a prey polypeptide which binds to a bait polypeptide of interest. The method in which protein-protein interactions are identified comprises the following steps: i) mating at least one first haploid recombinant yeast cell clone from a recombinant yeast cell clone library that has been transformed with a plasmid containing the prey polynucleotide to be assayed with a second haploid recombinant yeast cell clone transformed with a plasmid containing a bait polynucleotide encoding for the bait polypeptide; ii) cultivating diploid cell clones obtained in step i) on a selective medium; and iii) selecting recombinant cell clones which grow on the selective medium. This method may further comprise the step of: iv) characterizing the prey polynucleotide contained in each recombinant cell clone which is selected in step iii).

In yet another embodiment of the present invention, in lieu of yeast, Escherichia coli is used in a bacterial two-hybrid system, which encompasses a similar principle to that described above for yeast, but does not involve mating for characterizing the prey polynucleotide.

In yet another embodiment of the present invention, mammalian cells and a method. similar to that described above for yeast for characterizing the prey polynucleotide are used.

By performing the yeast, bacterial or mammalian two-hybrid system it is possible to identify for one particular bait an interacting prey polypeptide. The prey polynucleotide that has been selected by testing the library of preys in a screen using the two-hybrid, two plus one hybrid methods and the like, encodes the polypeptide interacting with the protein of interest.

The present invention is also directed, in a general aspect, to a complex of polypeptides, polynucleotides encoding the polypeptides composed of a bait polypeptide or bait polynucleotide encoding the bait polypeptide and a prey polypeptide or prey polynucleotide encoding the prey polypeptide capable of interacting with the bait polypeptide of interest. These complexes are identified in Table I. In another aspect, the present invention relates to a complex of polynucleotides consisting of a first polynucleotide, or a fragment thereof, encoding a prey polypeptide that interacts with a bait polypeptide and a second polynucleotide or a fragment thereof. This fragment has at least 12 consecutive nucleotides, but can have between 12 and 500 consecutive nucleotides, or between 12 and 1 ,500 consecutive nucleotides or between 12 and 3,000 consecutive nucleotides.

In yet another embodiment, the present invention relates to an isolated complex of at least two polypeptides encoded by two polynucleotides wherein said two polypeptides are associated in the complex by affinity binding and are depicted in Table I.

In yet another embodiment, the present invention relates to an isolated complex comprising at least a polypeptide as described in column 1 of Table I and a polypeptide as described in column 2 of Table I. The present invention is not limited to these polypeptide complexes alone but also includes the isolated complex of the two polypeptides in which fragments and/or homologous polypeptides exhibit at least 95% sequence identity, as well as from 96% sequence identity to 99.999% sequence identity.

Also encompassed in another embodiment of the present invention is an isolated complex comprising a polypeptide, or a nucleotide coding for this polypeptide (column 1 , Table II) and a SID® polypeptide (column 2, table II), or a polynucleotide coding for this SID® polypeptide (column 3, Table II) interacting with the polypeptide.

Besides the isolated complexes described above, polynucleotide coding for a

Selected Interacting Domain (SID®) polypeptide or a variant thereof or any of the polynucleotide set forth in Table II can be inserted into an expression vector which contains the necessary elements for the transcription and translation of the inserted protein-coding sequence. Such transcription elements include a regulatory region and a promoter. Thus, the polynucleotide which may encode a marker compound of the present invention is operably linked to a promoter in the expression vector. The expression vector may also include a replication origin. A wide variety of host/expression vector combinations are employed in expressing the nucleic acids of the present invention. Useful expression vectors that can be used include, for example, segments of chromosomal, non-chromosomal and synthetic DNA sequences. Suitable vectors include, but are not limited to, derivatives of SV40 and pcDNA and known bacterial plasmids such as col El, pCR1 , pBR322, pMal-C2, pET, pGEX as described by Smith et a) [need cite 1988], pMB9 and derivatives thereof, plasmids such as RP4, phage DNAs such as the numerous derivatives of phage I such as NM989, as weii as other phage DNA such as M13 and filamentous single stranded phage DNA; yeast plasmids such as the 2 micron plasmid or derivatives of the 2m plasmid, as well as centomeric and integrative yeast shuttle vectors; vectors useful in eukaryotic cells such as vectors useful in insect or mammalian cells; vectors derived from combinations of plasmids and phage DNAs, such as plasmids that have been modified to employ phage DNA or the expression control sequences; and the like.

For example in a bacuiovirus expression system, both non-fusion transfer vectors, such as, but not limited to pVL941 (SamHI cloning site Summers, pVL1393 (SamHI, Smal, Xba\, EcoRI, Λ/ofl, Xmalll, Sglll and Psfl cloning sites; Invitrogen) pVL1392 (Sglll, Pst\, Not\, Xmalll, EcoRI, Xba/I, Smal and SamHI cloning site; Summers and Invitrogen) and pBlueBaclll (SamHI, BglW, Pst\, Λ/col and HindW\ cloning site, with blue/white recombinant screening, Invitrogen), and fusion transfer vectors such as, but not limited to, pAc700(BamHI and Kpπl cloning sites, in which the SamHI recognition site begins with the initiation codon; Summers), pAc701 and pAc70-2 (same as pAc700, with different reading frames), pAc360 (SamHI cloning site 36 base pairs downstream of a polyhedrin initiation codon; Invitrogen (195)) and pBlueBacHisA, B, C ( three different reading frames with SamHI, BglW, Pst\, Λ/col and HindW\ cloning site, an N-terminal peptide for ProBond purification and blue/white recombinant screening of plaques; Invitrogen (220) can be used.

Mammalian expression vectors contemplated for use in the invention include vectors with inducible promoters, such as the dihydrofolate reductase promoters, any expression vector with a DHFR expression cassette or a DHFR/methotrexate co-amplification vector such as pED (Psfl, Sa/I, Sbal, Smal and EcoRI cloning sites, with the vector expressing both the cloned gene and DHFR; Kaufman, 1991 ). Alternatively a glutamine synthetase/methionine sulfoximine co-amplification vector, such as pEE14 (HindW\, Xba/I, Smal, Sba\, EcoRI and Bc/I cloning sites in which the vector expresses glutamine synthetase and the cloned gene; Celltech). A vector that directs episomal expression under the control of the Epstein Barr Virus (EBV) or nuclear antigen (EBNA) can be used such as pREP4 (SamHI, Sfi\, Xho\, Notl, Nhel, HindlW, Nhe\, PvuW and Kpn\ cloning sites, constitutive RSV-LTR promoter, hygromycin selectable marker; Invitrogen) pCEP4 (SamHI, Sfil, Xho\, Not\, Nhe\, HindlW, ^' Nhel, PvuW and Kpn\ cloning sites, constitutive hCMV immediate early gene promoter, hygromycin selectable marker; Invitrogen), pMEP4 {Kpnϊ, Pvu\, Nhe\, HindW\, Notl, Xhol, Sfil, SamHI cloning sites, inducible methallothionein lla gene promoter, hygromycin selectable marker, Invitrogen), pREPδ (SamHI, Xhol, Notl, HindlW, Nhel and Kpnl cloning sites, RSV-LTR promoter, histidinol selectable marker; Invitrogen), pREP9 (Kpnl, Nhel, HindlW, Notl, Xhol, Sfil, SamHI cloning sites, RSV-LTR promoter, G418 selectable marker; Invitrogen), and pEBVHis (RSV-LTR promoter, hygromycin selectable marker, N-terminal peptide purifiable via ProBond resin and cleaved by enterokinase; Invitrogen).

Selectable mammalian expression vectors for use in the invention include, but are not limited to, pRc/CMV (Hindl , BstXl, Notl, Sbal and Apal cloning sites, G418 selection, Invitrogen), pRc/RSV (Hindll, Spel, BstXl, Notl, Xbal cloning sites, G418 selection, Invitrogen) and the like. Vaccinia virus mammalian expression vectors (see, for example Kaufman 1991 that can be used in the present invention include, but are not limited to, pSC11 (Smal cloning site, TK- and β-gal selection), pMJ601 (Sail, Smal, Afl , Naή, SspMII, SamHI, Apal, Nhel, SacW, Kpnl and HindlW cloning sites; TK- and β-gal selection), pTKgptFIS (EcoRI, Psfl, Salll, Accl, Hindll, Sbal, SamHI and Hpa cloning sites, TK or XPRT selection) and the like.

Yeast expression systems that can also be used in the present include, but are not limited to, the non-fusion pYES2 vector (Xbal, Sphl, Shol, Notl, GstXl, EcoRI, BstXl, SamHI, Sad, Kpnl and HindlW cloning sites, Invitrogen), the fusion pYESHisA, B, C (Xiba/I, Sphl, Shol, Notl, BstXl, EcoRI, SamHI, Sad, Kpnl and HindlW cloning sites, N-terminal peptide purified with ProBond resin and cleaved with enterokinase; Invitrogen), pRS vectors and the like. Consequently, mammalian and typically human cells, as well as bacterial, yeast, fungi, insect, nematode and plant cells an used in the present invention and may be transfected by the nucleic acid or recombinant vector as defined herein.

Examples of suitable cells include, but are not limited to, VERO cells, HELA cells such as ATCC No. CCL2, CHO cell lines such as ATCC No. CCL61 , COS cells such as COS-7 cells and ATCC No. CRL 1650 cells, W138, BHK, HepG2, 3T3 such as ATCC No. CRL6361 , A549, PC12, K562 cells, 293 cells, Sf9 cells such as ATCC No. CRL1711 and Cv1 cells such as ATCC No. CCL70.

Besides the specific isolated complexes, as described above, the present invention relates to and also encompasses SID® polynucleotides. As explained above, for each bait polypeptide, several prey polypeptides may be identified by comparing and selecting the intersection of every isolated fragment that are included in the same polypeptide. Thus the SID® polynucleotides of the present invention are represented by the shared nucleic acid sequences of uneven SEQ ID from n°1 to 547 (column 2, Table II) encoding the SID® polypeptides of even SEQ ID from n°2 to 548 (column 3 of Table II).

The present invention is not limited to the SID® sequences as described in the above paragraph, but also includes fragments of these sequences having at least 12 consecutive nucleic acids, between 12 and 100 consecutive nucleic acids and between 12 and 400 consecutive nucleic acids and between 12 and 800 consecutive nucleic acids, as well as variants thereof. The fragments or variants of the SID® sequences possess at least the same affinity of binding to its protein or polypeptide counterpart, against which it has been initially selected. Moreover this variant and/or fragments of the SID® sequences alternatively can have between 95% and 99.999% sequence identity to its protein or polypeptide counterpart.

According to the present invention the variants can be created by known mutagenesis techniques either in vitro or in vivo. Such a variant can be created such that it has altered binding characteristics with respect to the target protein and more specifically that the variant binds the target sequence with either higher or lower affinity.

Polynucleotides that are complementary to the above sequences which include the polynucleotides of the SID®'s, their fragments, variants and those that have specific sequence identity are also included in the present invention.

The polynucleotide encoding the SID® polypeptide, fragment or variant thereof can also be inserted into recombinant vectors which are described in detail above.

The present invention also relates to a composition comprising the above-mentioned recombinant vectors containing the SID® polynucleotides in Table II, fragments or variants thereof, as well as recombinant host cells transformed by the vectors. The recombinant host cells that can be used in the present invention were discussed in greater detail above.

The compositions comprising the recombinant vectors can contain physiological acceptable carriers such as diluents, adjuvants, excipients and any vehicle in which this composition can be delivered therapeutically and can include, but is are not limited to sterile liquids such as water and oils.

In yet another embodiment, the present invention relates to a method of selecting modulating compounds, as well as the modulating molecules or compounds themselves which may be used in a pharmaceutical composition. These modulating compounds may act as a cofactor, as an inhibitor, as antibodies, as tags, as a competitive inhibitor, as an activator or alternatively have agonistic or antagonistic activity on the protein-protein interactions. The activity of the modulating compound does not necessarily, for example, have to be 100% activation or inhibition. Indeed, even partial activation or inhibition can be achieved that is of pharmaceutical interest.

The modulating compound can be selected according to a method which comprises:

(a) cultivating a recombinant host cell with a modulating compound on a selective medium and a reporter gene the expression of which is toxic for said recombinant host cell wherein said recombinant host cell is transformed with two vectors:

(i) wherein said first vector comprises a polynucleotide encoding a first hybrid polypeptide having a DNA binding domain;

(ii)wherein said second vector comprises a polynucleotide encoding a second hybrid polypeptide having a transcriptional activating domain that activates said toxic reporter gene when the first and second hybrid polypeptides interact; (b) selecting said modulating compound which inhibits or permits the growth of said recombinant host cell.

Thus, the present invention relates to a modulating compound that inhibits the protein-protein interactions of a complex of two polypeptides as described in Table I. The present invention also relates to a modulating compound that activates the protein-protein interactions of a complex of two polypeptides as described in Table I.

In yet another embodiment, the present invention relates to a method of selecting a modulating compound, which modulating compound inhibits the interactions of two polypeptides as described in Table I. This method comprises:

(i) wherein said first vector comprises a polynucleotide encoding a first hybrid polypeptide having a first domain of an enzyme;

(ii) wherein said second vector comprises a polynucleotide encoding a second hybrid polypeptide having an enzymatic transcriptiona) activating domain that activates said toxic reporter gene when the first and second hybrid polypeptides interact; (b) selecting said modulating compound which inhibits or permits the growth of said recombinant host cell.

In the two methods described above any toxic reporter gene can be utilized including those reporter genes that can be used for negative selection including the URA3 gene, the CYH1 gene, the CYH2 gene and the like.

In yet another embodiment, the present invention provides a kit for screening a modulating compound. This kit comprises a recombinant host cell which comprises a reporter gene the expression of which is toxic for the recombinant host cell. The host cell is transformed with two vectors. The first vector comprises a polynucleotide encoding a first hybrid polypeptide having a DNA binding domain; and the second vector comprises a polynucleotide encoding a second hybrid polypeptide having a transcriptional activating domain that activates said toxic reporter gene when the first and second hybrid polypeptides interact.

In yet another embodiment a kit is provided for screening a modulating compound by providing a recombinant host cell, as described in the paragraph above, but instead of a DNA binding domain, the first vector encodes a first hybrid polypeptide containing a first domain of a protein. The second vector encodes a second polypeptide containing a second part of a complementary domain of a protein that activates the toxic reporter gene when the first and second hybrid polypeptides interact.

In the selection methods described above, the activating domain can be p42 Gal 4, YP16 (HSV) and the DNA-binding domain can be derived from Gal4 or Lex A. The protein or enzyme can be adenylate cyclase, guanylate cyclase, DHFR and the like.

Examples of modulating compounds are set forth in column 2 and 3 of Table II.

In yet another embodiment, the present invention relates to a pharmaceutical composition comprising the modulating compounds for preventing or treating Candida infection, cancer and neurodegenerative diseases in a human or animal, most preferably in a mammal.

This pharmaceutical composition comprises a pharmaceutically acceptable amount of the modulating compound. The pharmaceutically acceptable amount can be estimated from cell culture assays. For example, a dose can be formulated in animal models to achieve a circulating concentration range that includes or encompasses a concentration point or range having the desired effect in an in vitro system. This information can thus be used to accurately determine the doses in other mammals, including humans and animals.

The therapeutically effective dose refers to that amount of the compound that results in amelioration of symptoms in a patient. Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or in experimental animals. For example, the LD50 (the dose lethal to 50% of the population) as well as the ED50 (the dose therapeutically effective in 50% of the population) can be determined using methods known in the art. The dose ratio between toxic and therapeutic effects is the therapeutic index which can be expressed as the ratio between LD 50 and ED50 compounds that exhibit high therapeutic indexes.

The data obtained from the cell culture and animal studies can be used in formulating a range of dosage of such compounds which lies preferably within a range of circulating concentrations that include the ED50 with little or no toxicity.

The pharmaceutical composition can be administered via any route such as locally, orally, systemically, intravenously, intramuscularly, mucosally, using a patch and can be encapsulated in liposomes, microparticles, microcapsules, and the like. The pharmaceutical composition can be embedded in liposomes or even encapsulated.

Any pharmaceutically acceptable carrier or adjuvant can be used in the pharmaceutical composition. The modulating compound will be preferably in a soluble form combined with a pharmaceutically acceptable carrier. The techniques for formulating and administering these compounds can be found in "Remington's Pharmaceutical

Sciences" Mack Publication Co., Easton, PA, latest edition. The mode of administration optimum dosages and galenic forms can be determined by the criteria known in the art taken into account the seriousness of the general condition of the mammal, the tolerance of the treatment and the side effects.

The present invention also relates to a method of treating or preventing Candida infection, cancer and neurodegenerative diseases in a human or mammal in need of such treatment. This method comprises administering to a mammal in need of such treatment a pharmaceutically effective amount of a modulating compound which binds to a targeted Candida protein or a protein involved in cancer or neurodegenerative diseases. In a preferred embodiment, the modulating compound is a polynucleotide which may be placed under the control of a regulatory sequence which is functional in the mammal or human.

In yet another embodiment, the present invention relates to a pharmaceutical composition comprising a SID® polypeptide, a fragment or variant thereof. The SID® polypeptide, fragment or variant thereof can be used in a pharmaceutical composition provided that it is endowed with highly specific binding properties to a bait polypeptide of interest.

The original properties of the SID® polypeptide or variants thereof interfere with the naturally occurring interaction between a first, protein and a second protein within the cells of the organism. Thus, the SID® polypeptide binds specifically to either the first polypeptide or the second polypeptide.

Therefore, the SID® polypeptides of the present invention or variants thereof interfere with protein-protein interactions between Candida proteins or between an Candida proteins and a human or mammal protein.

Thus, the present invention relates to a pharmaceutical composition comprising a pharmaceutically acceptable amount of a SID® polypeptide or variant thereof, provided that the variant has the above-mentioned two characteristics; i.e., that it is endowed with highly specific binding properties to a bait polypeptide of interest and is devoid of biological activity of the naturally occurring protein. In yet another embodiment, the present invention relates to a pharmaceutical composition comprising a pharmaceutically effective amount of a polynucleotide encoding a SID® polypeptide or a variant thereof wherein the polynucleotide is placed under the control of an appropriate regulatory sequence. Appropriate regulatory sequences that are used are polynucleotide sequences derived from promoter elements and the like.

Polynucleotides that can be used in the pharmaceutical composition of the present invention include the nucleotide sequences of uneven SEQ ID from n°1 to 547 (column 2, Table II).

Besides the SID® polypeptides and polynucleotides, the pharmaceutical composition of the present invention can also include a recombinant expression vector comprising the polynucleotide encoding the SID® polypeptide, fragment or variant thereof.

The above described pharmaceutical compositions can be administered by any route such as orally, systemically, intravenously, intramuscularly, intradermally, mucosally, encapsulated, using a patch and the like. Any pharmaceutically acceptable carrier or adjuvant can be used in this pharmaceutical composition.

The SID® polypeptides as active ingredients will be preferably in a soluble form combined with a pharmaceutically acceptable carrier. The techniques for formulating and administering these compounds can be found in "Remington's Pharmaceutical Sciences" supra.

The amount of pharmaceutically acceptable SID® polypeptides can be determined as described above for the modulating compounds using cell culture and animal models.

Such compounds can be used in a pharmaceutical composition to treat or prevent Candida infection, cancer and neurodegenerative diseases.

Thus, the present invention also relates to a method of preventing or treating Candida infection, cancer and neurodegenerative diseases in a mammal said method comprising the steps of administering to a mammal in need of such treatment a pharmaceutically effective amount of:

(1) a SID® polypeptide of even SEQ ID from n°2 to 548 (column 3, Table II) or a variant thereof which binds to a targeted Candida, yeast, human or mammal protein; or

(2) or SID® polynucleotide encoding a SID® polypeptide of SEQ ID from n°2 to 548 (column 3, Table II) or a variant or a fragment thereof wherein said polynucleotide is placed under the control of a regulatory sequence which is functional in said mammal; or

(3) a recombinant expression vector comprising a polynucleotide encoding a SID® polypeptide which binds to a yeast, Candida protein.

In another embodiment the present invention nucleic acids comprising a sequence of uneven SEQ ID from n°2 to 548 (column 2, Table II) which encodes the protein of even SEQ ID from n°2 to 548 (column 3, Table ll)and/or functional derivatives thereof are administered to modulate complex (from Table I) function by way of gene therapy. Any of the methodologies relating to gene therapy available within the art may be used in the practice of the present invention such as those described by Goldspiel et al Clin. Pharm. 12 pgs. 488-505 (1993).

Delivery of the therapeutic nucleic acid into a patient may be direct in vivo gene therapy (i.e., the patient is directly exposed to the nucleic acid or nucleic acid-containing vector) or indirect ex vivo gene therapy (i.e., cells are first transformed with the nucleic acid in vitro and then transplanted into the patient).

For example for in vivo gene therapy, an expression vector containing the nucleic acid is administered in such a manner that it becomes intracellular; i.e., by infection using a defective or attenuated retroviral or other viral vectors as described, for example in U.S. Patent 4,980,286 or by Robbins et al, Pharmacol. Ther. , 80 No. 1 pgs. 35-47 (1998).

The various retroviral vectors that are known in the art are such as those described in Miller et al, Meth. Enzymol. 217 pgs. 581-599 (1993) which have been modified to delete those retroviral sequences which are not required for packaging of the viral genome and subsequent integration into host cell DNA. Also adenoviral vectors can be used which are advantageous due to their ability to infect non-dividing cells and such high-capacity adenoviral vectors are described in Kochanek, Human Gene Therapy, 10, pgs. 2451-2459 (1999). Chimeric viral vectors that can be used are those described by Reynolds et al, Molecular Medecine Today, pgs. 25 -31 (1999). Hybrid vectors can also be used and are described by Jacoby et al, Gene Therapy, 4, pgs. 1282-1283 (1997).

Direct injection of naked DNA or through the use of microparticle bombardment (e.g., Gene Gun®; Biolistic, Dupont). or by coating it with lipids can also be used in gene therapy. Cell-surface receptors/transfecting agents or through encapsulation in liposomes, microparticles or microcapsules or by administering the nucleic acid in linkage to a peptide which is known to enter the nucleus or by administering it in linkage to a ligand predisposed to receptor-mediated endocytosis ( See, Wu & Wu, J. Biol. Chem., 262 pgs. 4429-4432 ( 1987)) can be used to target cell types which specifically express the receptors of interest.

In another embodiment a nucleic acid ligand compound may be produced in which the ligand comprises a fusogenic viral peptide designed so as to disrupt endosomes, thus allowing the nucleic acid to avoid subsequent lysosomal degradation. The nucleic acid may be targeted in vivo for cell specific endocytosis and expression by targeting a specific receptor such as that described in WO92/06180, WO93/14188 and WO 93/20221. Alternatively the nucleic acid may be introduced intracellularly and incorporated within the host cell genome for expression by homologous recombination. See, Zijlstra et al, Nature, 342, pgs. 435-428 (1989).

In ex vivo gene a gene is transferred into cells in vitro using tissue culture and the cells are delivered to the patient by various methods such as injecting subcutaneously, application of the cells into a skin graft and the intravenous injection of recombinant blood cells such as hematopoietic stem or progenitor cells.

Cells into which a nucleic acid can be introduced for the purposes of gene therapy include, for example, epithelial cells, endotheiial cells, keratinocytes, fibroblasts, muscle cells, hepatocytes and blood cells. The blood cells that can be used include, for example,

T-lymphocytes, B-lymphocytes, monocytes, macrophages, neutrophils, eosinophils, megakaryotcytes, granulocytes, hematopoietic cells or progenitor cells and the like. In yet another embodiment the present invention relates to protein chips or protein microarrays. It is well known in the art that microarrays can contain more than 10,000 spots of a protein that can be robotically deposited on a surface of a glass slide or nylon filter. The proteins attach covalently to the slide surface, yet retain their ability to interact with other proteins or small molecules in solution. In some instances the protein samples can be made to adhere to glass slides by coating the slides with an aldehyde-containing reagent that attaches to primary amines. A process for creating microarrays is described, for example by MacBeath and Schreiber in Science, Volume 289, Number 5485, pgs, 1760-1763 (2000) or Service, Science, Vol, 289, Number 5485 pg. 1673 (2000). An apparatus for controlling, dispensing and measuring small quantities of fluid is described, for example, in U.S. Patent No. 6,1 12,605.

The present invention also provides a record of protein-protein interactions, PIM®'s, SID®'s and any data encompassed in the following Tables. It will be appreciated that this record can be provided in paper or electronic or digital form.

In order to fully illustrate the present invention and advantages thereof, the following specific examples are given, it being understood that the same are intended only as illustrative and in nowise limitative.

EXAMPLES

Example 1 : Preparation of a Saccharomyces cerevisiae genomic collection 1 .A. Collection preparation and transformation in E. coli 1 .B. Collection transformation in Saccharomyces cerevisiae 1 .C. Construction of bait plasmid Example 2: Screening the collection with the two-hybrid in yeast system 2. A. The mating protocol

2.B. The X-Gal overlay assay 2.C. The luminometry assay Example 3: Identification of positive clones 3.A. PCR on yeast colonies

3.B. Plasmid rescue from yeast by electroporation Example 4: Protein-protein interaction Example 5: Identification of SID® Example 6: Screening of modulating agent

Example 7: Making of polyclonal and monoclonal antibodies

Medium composition and standard protocols are available in Maniatis et al..

Example 1 : Preparation of a Saccharomyces cerevisiae genomic collection

1.A. Collection preparation and transformation in Escherichia coli

1.A.1. Fragmented of genomic DNA preparation The Saccharomyces cerevisiae (strain YM955 (Matα, ura3-52, his3-200, ade2-101 ,

Iys2-801 , leu2-3, trp1-901 , tyr1-501 , gal4-542, gal80-538)) genomic DNA was fragmented in a nebulizer (GATC) for 1 minute, precipitated and resuspended in water.

The obtained nebulized genomic DNA was successively treated with Mung Bean Nuclease (Biolabs) (30 minutes at 30°C), T4 DNA polymerase (Biolabs) (10 minutes at 37°C) and Klenow enzyme (Pharmacia) (10 minutes at room temperature and 1 hour at 16°C).

DNA was then extracted, precipitated and resuspended in water.

1.A.2. Ligation of linkers to blunt-ended genomic DNA Oligonucleotide PL160 (5' end phosphorylated) 1 μg/μl and PL159 2μg/μl were used.

Sequence of the oligo PL160 : 5'-ATCCCGGACGAAGGCC-3' (SEQ ID No. 549) Sequence of the oligo PL159 : 5'-GGCCTTCGTCCGG-3' (SEQ ID No. 550)

Linkers were preincubated (5 minutes at 95°C, 10 minutes at 68°C, 15 minutes at 42°C) then cooled down at room temperature and ligated with genomic DNA inserts at 4°C overnight.

Linkers were further removed on a separation column (Chromaspin TE 400, Clontech), according to the manufacturer protocol. 1.A.3. Vector preparation pP2 (Figure 8) was successively digested with SamHI restriction enzyme (Biolabs) for 1 hour at 37°C, dephosphorylated with Calf Intestine Phosphatase (CIP) (Biolabs) and filled in with dGTP using Vent DNA polymerase (exo-) (Biolabs), extracted, precipitated and resuspended in water.

1.A.4. Ligation between vector and insert of genomic DNA

The prepared vector was ligated overnight at 15°C with the genomic blunt ended DNA described in section 2 using T4 DNA ligase (Biolabs). The DNA was then precipitated and resuspended in water.

1.A.5. Library transformation in Escherichia coli

DNA from section 1 .A.4 was then transformed into Electromax DH10B eiectrocompetent cells (Gibco BRL) with Cell Porator apparatus (Gibco BRL). 1 ml SOC medium was added and transformed cells were incubated at 37°C for 1 hour. 9 ml volumes of SOC medium per tube were added and plated on LB+ampicillin medium.

Colonies were scraped with liquid LB medium, aliquoted and frozen at -80°C.

The obtained collection of recombinant cell clones is named HGXYeastB.

1.B. Collection transformation in Saccharomyces cerevisiae

The Saccharomyces cerevisiae strain (Y187 (MATα Gal4Δ GalδOΔ ade2-101 His3 Leu2-3, -1 12 Trp1-901 Ura3-52 URA3::UASGAL1 -LacZ Met)) was transformed with the HGXYeastB Saccharomyces cerevisiae genomic DNA library.

The plasmid DNA contained in E. coli was extracted (Qiagen) from aliquoted E. coli frozen cells (1.A.5.).

Saccharomyces cerevisiae yeast Y187 was grown in YPGIu. Yeast transformation was performed according to standard protocol (Giest et al. Yeast, 11 , 355-360, 1995) using yeast carrier DNA (Clontech). This experiment lead to 10⁴ to 5.10⁴ cells/μg DNA. 2.10⁴ cells were spread on DO-Leu medium per plates. Cells were aliquoted and frozen at -80°C.

The obtained collection of recombinant cell clones is named HGXYeastY. 1.C. Construction of bait plasmid

The genomic amplification of the ORF was obtained by PCR using the Pfu proofreading Tag polymerase (Stratagene) and 200 ng of genomic DNA as template. PCR primers were chosen in regions flanking the ORF. PCR program was set up as followed :

94° 45"

48° 45" x 30 cycles

72° 6'

72° 10'

15° oo

The amplification was checked on agarose gel.

PCR fragments were purified with Qiaquick column (Qiagen) according to the manufacturer protocol.

Purified PCR fragments were digested with adequate restriction enzymes.

Digested PCR fragments were purified with Qiaquick column (Qiagen) according to the manufacturer protocol.

Digested and purified PCR fragments were ligated into an adequately digested and dephosphorylated bait vector (pB6, Figure 3) according to standard protocol (Maniatis et al.).

Competent bacterial cells were transformed. Cells were grown, DNA was extracted and plasmid was sequenced.

Example 2: Screening the collection with the two-hybrid in yeast system

2.A. The mating protocol The mating two-hybrid in yeast system has been chosen (firstly described by Legrain et al., Nature Genetics, 1997, vol. 16, 277-282, Toward a functional analysis ofthe yeast genome through exhaustive two-hybrid screens) for its advantages but the Saccharomyces cerevisiae collection could also have been screened in classical two- hybrid system as described in Fields et al. or in a yeast reverse two-hybrid system. The mating procedure allows a direct selection on selective plates because the two fusion proteins are already produced in the parental cells. No replica plating is required.

This protocol was written for the use of the library transformed into the Y187 strain.

Before mating, S. cerevisiae (CG 1945 strain (MATa Gal4-542 Gah 80-538 ade2- 101 His3*200 Leu2-3,-112 Trp1-901 Ura3-52 Lys2-801 URA3::GAL4 17mers (X3)- CyC1TATA-LacZ LYS2::GAL1 UAS-GAL1TATA-HIS3 CYH^R)) was transformed according to step 1.B. and spread on DO-Trp medium.

Day 1 , morning : preculture Y187 cells carrying the bait plasmid obtained at step 1.C. were precultured in 20 ml DO- Trp medium and grown at 30°C with vigorous agitation.

Day 1, late afternoon : culture

The OD_600nm of the DO-Trp preculture of Y187 cells carrying the bait plasmid was measured. The OD₆₀onm must lie between 0.1 and 0.5 in order to correspond to a linear measurement.

150 ml DO-Trp at OD600nm 0.006/ml were inoculated and grown overnight at 30°C with vigorous agitation.

Day 2 : mating medium and plates

5 YPGIu plates

50 ml tube with 30 ml DO-Leu-Trp-His

100 ml flask with 20 ml of YPGIu 75 DO-Leu-Trp-His plates

2 DO-Leu plates

2 DO-Trp plates

2 DO-Leu-Trp plates

The OD600nm of the DO-Trp culture was measured. It should be around 1. For the mating, twice as many bait cells as library cells were used. To get a good mating efficiency, one must collect the cells at 10⁸ cells per cm².

The amount of bait culture (in ml) that makes up 80 OD600nm units for the mating with the yeast collection was estimated. A vial containing the HGXYeastY library was thawed slowly on ice. The contents of the vial were added to 20 ml YPGIu. Cells were recovered at 30°C, under gentle agitation for

10 minutes.

Mating The 80 OD600nm units of bait culture was placed into a 250 ml flask

The HGXYeastY library culture was added to the bait culture. The mixture of diploids was transferred into 50 ml sterile tubes and centrifuged. The supernatant was discarded and the cells resuspended in YPGIu medium.

Cells were distributed in 400 μl samples in YPGIu plates with glass beads and spread by shaking the plates.

Plates were incubated cells-up at 30°C for 4h30min.

Collection of mated cells

Plates were washed and rinsed, and collected cells were spread on DO-Leu-Trp-His+Tet plates.

Day 4

Clones that were able to grow on DO-Leu-Trp-His+Tetracyclin were selected. This medium allows one to isolate diploid clones presenting an interaction. The His+ colonies were counted on control plates. The number of His+ cell clones will define which protocol is to be processed: Upon 20.10⁶ His+ colonies :

- if number of His+ cell clones > 285 : then process overlay and then luminometry protocols on blue colonies (2.B and 2.C).

- if number of His+ cell clones < 285 : process luminometry protocol (2.C). The following step leads to the selection of the strongest interaction.

2.B. The X-Gal overlay assay X-Gal overlay assay was performed directly on the selective medium plates after scoring the number of His⁺ colonies. Materials

A waterbath was set up. The water temperature should be 50°C. . 0.5 M Na₂HPO₄ pH 7.5. • 1.2% Bacto-agar. • 2% X-Gal in DMF.

• Overlay mixture : 0.25 M Na₂HPO₄ pH7.5, 0.5% agar, 0.1% SDS, 7% DMF (LABOSI), 0.04% X-Gal (ICN). For each plate, 10 ml overlay mixture are needed.

• DO-leu-trp-his plates. • Sterile toothpicks.

Experiment

Temperature of the overlay mix should be between 45 and 50°C.

The overlay-mix was poured over the plates in portions of 10 ml and collected when the top layer was settled. Plates were incubated overlay-up at 30°C. The time was noted.

Blue colonies were regularly checked for. If no blue colony appeared, overnight incubation was performed. Using a pen the number of positives was marked.

The positives colonies were streaked on fresh DO-Leu-Trp-His plates with a sterile toothpick.

2.C. The luminometry assay His+ colonies were grown overnight at 30°C in microtiter plates containing DO-Leu-Trp- His+Tetracyclin medium with shaking. The day after, the overnight culture was diluted 15 times into a new microtiter plate containing the same medium, and incubated 5 hours at 30°C with shaking. Samples were diluted 5 times and the OD₆oon was read. Another dilution was performed to obtain between 10,000 and 75,000 yeast cells/well in 100 μl final volume.

Per well, 76 μl of One Step Yeast Lysis Buffer (Tropix), 20 μl Sapphirell Enhancer (Tropix), 4 μl Galacton Star (Tropix) were added, and incubated 40 minutes at 30°C. The β-Gal read-out (L) was measured using a Luminometer (Trilux, Wallach).

The value of ODeoonm L was calculated and interacting preys having highest values were selected.

At this step of the protocol, diploid cell clones presenting interaction were isolated. The next step was aimed at identifying polypeptides involved in the selected interactions.

Example 3: Identification of positive clones 3. A. PCR on yeast colonies

Introduction

PCR amplification of fragments of plasmid DNA directly on yeast colonies was a quick and efficient procedure to identify sequences cloned into this plasmid. It is directly derived from a published protocol (Wang H. et al., Analytical Biochemestry, 237, 145-146, 1996). However, it is not a standardized protocol: it varies from strain to strain and it is dependent of experimental conditions (number of cells, Taq polymerase source, etc). This protocol should be optimized to specific local conditions.

Materials

- For 1 well, PCR mix composition was : 32.5 μl water,

5 μl 10X PCR buffer (Pharmacia), 1 μl dNTP 10 mM,

0.5 μl Taq polymerase (5u/μl) (Pharmacia),

0.5 μl oligonucleotide ABS1 10 pmole/μl: 5'-GCGTTTGGAATCACTACAGG-3' (SEQ ID No. 551 )

0.5 μl oligonucleotide ABS2 10 pmole/μl: 5 -CACGATGCACGTTGAAGTG-3' (SEQ ID No. 552).

- 1 N NaOH.

Experiment

Positive colonies were grown overnight at 30°C on a 96 well cell culture cluster (Costar), containing 150 μl DO-Leu-Trp-His+Tetracyclin with shaking. Cultures were resuspended.

100 μl were transferred immediately on a Thermowell 96 (Costar) and centrifuged

5 minutes at 4000 rpm at room temperature.

Supernatant was removed. 5 μl NaOH were added to each well and shaken 1 minute.

The Thermowell was placed in the thermocycler (GeneAmp 9700, Perkin Elmer) 5 minutes at 99.9°C and then 10 minutes at 4°C.

In each well, the PCR mix was added and shaken well.

The PCR program was set up as followed : 94°C 3 minutes

94°C 30 secondes

53°C 1 minute 30 secondes x 35 cycles

72°C 3 minutes

72°C 5 minutes

15°C oo

The quality, the quantity and the length of the PCR fragment was checked on agarose gel. The length of the cloned fragment was the estimated length of the PCR fragment minus 300 base pairs that correspond to the amplified flanking plasmid sequences.

3.B. Plasmid rescue from yeast by electroporation

Introduction The previous protocol of PCR on yeast cell may not be successful, in such a case, plasmids may be rescued from yeast by electroporation. This experiment allows the recovery of prey plasmids from yeast cells by transformation of £. coli with a yeast cellular extract. The prey plasmid can then be amplified and the cloned fragment sequenced.

Material

Plasmid rescue

Glass beads 425-600 μm (Sigma)

Phenol/chloroform (1/1) premixed with isoamyl alcohol (Amresco)

Extraction buffer: 2% Triton X100, 1 % SDS, 100 mM NaCl, 10 mM TrisHCI pH 8.0, 1 mM EDTA pH 8.0.

Mix ethanol/NH Ac: 6 volumes ethanol with 7.5 M NH Acetate, 70% Ethanol and yeast cells in patches on plates. Electroporation

SOC medium M9 medium

Selective plates : M9-Leu+Ampicillin

2 mm electroporation cuvettes (Eurogentech) Experiment

Plasmid rescue

The cell patch was prepared on DO-Leu-Trp-His with cell culture of section 2.C..

The cell of each patch was scraped in Eppendorf tube, 300 μl of glass beads were added in each tube, then, 200 μl extraction buffer and 200 μl phenol:chloroform:isoamyl alcohol

(25:24:1 ) were added.

Tubes were centrifuged 10 minutes at 15000 rpm.

180 μl supernatant were transferred to a sterile Eppendorf tube and 500 μl ethanol/NH Ac were added. Tubes were vortexed, centrifuged 15 minutes at 15000 rpm at 4°C.

The pellet was washed with 200 μl 70% ethanol, ethanol was removed and the pellet was dried and resuspend in 10 μl water. Extracts were stored at -20°C. Electroporation

Material: Eiectrocompetent MC1066 cells prepared according to standard protocols (Maniatis).

1 μl of yeast plasmid DNA-extract was added to pre-chilled Eppendorf tube, and kept on ice.

1 μl plasmid yeast DNA-extract sample was mixed and 20 μl eiectrocompetent cells were added and transferred in a cold electroporation cuvette. The Biorad eiectroporator was set on 200 ohms resistance, 25 μF capacity; 2.5 kVolts.

The cuvette was placed in the cuvette holder and electroporation was performed.

1 ml SOC was added into the cuvette and the cell-mix was transferred into sterile

Eppendorf tube.

Cells were recovered for 30 minutes at 37°C, spun down 1 minute, 4000x g and the supernatant was poured off. About 100 μl medium was kept and used to resuspend the cells. Cells were spread on selective plates (e.g., M9-Leu plates).

Plates were incubated for 36 hours at 37°C.

One colony was grown and plasmids were extracted. The presence and the size of the insert were checked through enzymatic digestion and agarose gel. The insert was sequenced.

Example 4: Protein-protein interaction For each bait, the previously protocol leads to the identification of prey polynucleotide sequences. In order to identify a protein-protein interaction, the obtained prey polypeptide sequence has to be characterized regarding the Saccharomyces cerevisiae genome. This was accomplished with a software program names blastwun (available on the

Internet site if the University of Washington : http://bioweb.pasteur.fr/seqanal/interfaces/blastwu.html, this is a development version of software for gene and protein identification through similarity searches of protein and nucleotide sequence databases). Blastwun program compares prey polynucleotide insert sequence (rescued from prey plasmid) with whole Saccharomyces cerevisiae genome (available on Stanford web site: http://qenome-www.stanford.edu/Saccharomyces/). This comparison lead to prey polynucleotide localizations in S. cerevisiae genome, each localization having a score depending on the homology of sequence. For each prey polynucleotide, the localization with the highest score was considered and, if the insert sequence was included in and was in phase with an Open Reading Frame, one prey polypeptide interacting with one bait polypeptide was identified. See Table I : Protein-protein interactions in Saccharomyces cerevisiae.

Example 5 : Identification of SID®

By comparing and selecting the intersection of every isolated prey fragments obtained from example 3 and that were included in the same polypeptide, one can define the Selected Interacting Domain (SID®) as illustrated in Figure 15. The SID® were illustrated in Table II.

Example 6: Screening of modulating agent

One specific interaction is selected.

A permeabilized yeast cell is transformed with plasmids containing bait polypeptide and prey polypeptide of the specific interaction. A top agar is plated containing transformed permeabilized yeast cells on square boxes (that already contains agarose gel).

The compounds to test are applied by spotting on top agar as soon as it is solidified, and incubated overnight at 30°C. Results are analyzed: lead compounds that prevent transformed permeabilized yeast cells from growing are selected.

Example 7: Making of polyclonal and monoclonal antibodies

The protein-protein complex of Table 1 was injected into mice and polyclonal and monoclonal antibodies were made following the procedure set forth in Sambrook et al supra.

More specifically mice are immunized with an immunogen comprising complexes conjugated to keyhole limpet hemocyanin using glutaraldehyde or EDC as is well known in the art. The complexes can also be stabilized by crosslinking as described in WO 00/37483. The immunogen is then mixed with an adjuvant. Each mouse receives four injections of 10 μg to 100 μg of immunogen, and after the fourth injection, blood samples are taken from the mice to determine if the serum contains antibodies to the immunogen. Serum titer is determined by ELISA or RIA. Mice with sera indicating the presence of antibody to the immunogen are selected for hybridoma production.

Spleens are removed from immune mice and single-cell suspension is prepared (Harlow et al 1988). Cell fusions are performed essentially as described by Kohler et al.. Briefly, P365.3 myeloma cells (ATTC Rockville, Md) or NS-1 myeloma cells are fused with spleen cells using polyethylene glycol as described by Harlow et al. Cells are plated at a density of 2 x 10⁵ cells/well in 96-well tissue culture plates. Individual wells are examined for growth and the supematants of wells with growth are tested for the presence of Table I complex-specific antibodies by ELISA or RIA using Table I complex as a target protein. Cells in positive wells are expanded and subcloned to establish and confirm monoclonality. Clones with the desired specificities are expanded and grown as ascites in mice or in a hollow fiber system to produce sufficient quantities of antibodies for characterization and assay development. Antibodies are tested for binding to bait polypeptide (from column 1 of Table I) aione or to prey polypeptide (from column 2 of Table I) alone, to determine which are specific for the Table I complex as opposed to those that bind to the individual proteins.

Monoclonal antibodies against each of the complexes set forth in Table I are prepared in a similar manner by mixing specified proteins together, immunizing an animal, fusing spleen cells with myeloma cells and isolating clones which produce antibodies specific for the protein complex, but not for individual proteins.

The following results obtained from these Examples, as well as the teachings in the specification are set forth in the Tables I and II below.

Table I depicts Saccharomyces cerevisiae interacting proteins. Table II provides polynucleotide (column 2) and polypeptide (column 3) sequences of SID® interacting with a given bait (column 1 ).

All the non-patented websites cited in the present specification are incorporated herein by reference.

While the invention has been described in terms of the various preferred embodiments, the skilled artisan will appreciate that various modifications, substitutions, omissions and changes may be made without departing from the scope thereof.

Accordingly, it is intended that the present invention be limited by the scope of the following claims, including equivalents thereof. Table I - Complexes of Interacting Proteins

Table II - SID® sequences interacting with a given bait

YDL10 |11 GCGGCAGAACGACAGGCAGCCATACTATCAAATGGACCCTCACCAATCACCAG 12 RQNDRQPYYQMDPHQS 5w CTGATAATGCCGCATCGCCTACGAAGAGCGTGAAGGCAACCACTAAAAATTCG PADNAASPTKSVKATTK

TCCACGAATAATAATGTCAATAGCAACAACAGCAATAATAACAGTAACCATGAT NSSTNNNVNSNNSNNNS

ATACTGAATTTTAATGATAACTATACTACCATTCTGCAACATTTGGCAAACGACC NHDILNFNDNYTTILQHL

ATCCTAATATACTGAGGGAGAAAGGAGGATCACAACAACAACAGCATCAGCAG ANDHPNILREKGGSQQQ

CAGCAACAACAGCAACAACAGCAGCAACAACAGCAGCAACAACAGAGCCTGG QHQQQQQQQQQQQQQ

ATACCCTTTTGCACCATTATCAAAGTTTACTCTCCAAGAGTGATAATGCAATAGCl QQQQSLDTLLHHYQSLL

CTTTGATGACAATGTTAGTAACAGCGCAGATCATAATGGCAGTAATAGCAACAA SKSDNAIAFDDNVSNSA

TAATAACAACAATAATGACATATCTAGTCCCGGTAATCTGATGGGATCTTGCAA DHNGSNSNNNNNNNDIS

TCAATGTAGATTAAAGAAAACAAAGTGCAACTATTTTCCCGACCTAGGCAACTG SPGNLMGSCNQCRLKKT

TCTCGAATGTGAAACGTCAAG KCNYFPDLGNCLECETS

YDL10 13 GCCGACCAAGTCAACTTTTAGTCGCTGGAAGAAGGCGGACCTAATTGACCTGG 14 PTKSTFSRWKKADLIDLA 5w CCAACAAGCTGGAAATTGACGGCTTCCCCAATTATGCCAAGAAGAGTGATATG NKLEIDGFPNYAKKSDMI

ATCGATTACCTCGAATCGCACTTGAATCATCTTGAGAAACCTGTGGATTTTAAA DYLESHLNHLEKPVDFK

GACGACTACCCGGAACTAAGGTCCTTTTATGAGTCGATGACAGTGGACCAGTC DDYPELRSFYESMTVDQ

AAAGGATGAACGGAACGAATACGGATCCGGATCTGGAAACGGGTCTGGGTCT SKDERNEYGSGSGNGS

GGATCCTGTGACACCGCCACAAATGATTCCGATCTGGAGAAAGCGTACATCAA GSGSCDTATNDSDLEKA

GGAGGATGATGATGAGAAACCGCAATCAGGCGATGAAACAAGCGCGACAAAG YIKEDDDEKPQSGDETS

CCGCTTTCCAGCAGGAATGCAAATTCAAACGCCAAAACGAACTTCAATTTGTTG ATKPLSSRNANSNAKTN

GATTTTAGCACAGATAACGACTCGTCCACCTCTGCGTTTACTAAGTTCAAGTTT FNLLDFSTDNDSSTSAFT

AACTTTCAGGAATACCTCTCCGATATTAGATACCAAACGCAGAAATTGAATGAA KFKFNFQEYLSDIRYQTQ

AATGTTCAGGACTATCTTTCCACTATTTCCGCCGTCGATACGATTTTTTCCTTGT KLNENVQDYLSTISAVDT

TGGAATTCTCTTTTCTGGTGAGAAACATATTAGCCGCCGGGCAGCCAACTTCTT IFSLLEFSFLVRNILAAGQ

CTTCTTCGCTCGCATCGTCGCTGGAAGCCGCCGTTGCCG PTSSSSLASSLEAAVA

ITCTA DKGLQTDDDEDWSTKA KKKKGKQPKKNSKSTKS TPSLSTLPSSMSPTSAIE VCTTCGESF

YDL10 29 TCGCAATTTCAGACACAACAGAACCGTGACATCAGCACGATGATGGAACACAC 30 SQFQTQQNRDISTMMEH 5w TAACAGTAATGATATGAGCGGCTCTGGTAAAAATCTCAAGAAACGTGTATCAAA TNSNDMSGSGKNLKKRV

GGCCTGTGACCATTGCCGAAAAAGAAAAATCAGATGTGATGAAGTGGACCAGC SKACDHCRKRKIRCDEV

AGACCAAAAAATGTTCAAACTGTATTAAGTTTCAGTTGCCTTGCACTTTCAAACA DQQTKKCSNCIKFQLPC

TCGTGATGAGATTCTTAAGAAGAAAAGAAAATTAGAAATCAAACATCATGCAAC TFKHRDEILKKKRKLEIKH

ACCAGGGGAATCACTTCAAACCTCAAATAGTATTAGCAATCCTGTAGCGTCTTC HATPGESLQTSNSISNPV

TTCAGTACCGAACAGTGGAAGGTTTGAACTTTTAAACGGTAATTCCCCCTTAGA ASSSVPNSGRFELLNGN

AAGCAATATCATCGATAAAGTCTCCAATATTCAAAATAATCTTAACAAAAAAATG SPLESNIIDKVSNIQNNLN

AATTCAAAGATTGAAAAATTGGATAGAAAAATGTCTTACATTATTGACAGTGTGG KKMNSKIEKLDRKMSYII

CTAGACTTGAGTGGTTATTGGACAAGGCTGTTAAAAAGCAGGAAGGCAAATAC DSVARLEWLLDKAVKKQ

AAGGAAAAGAACAATTTGCCCAAACCAGCGAGAAAAATATACTCTACGGCACTT EGKYKEKNNLPKPARKIY

TTAACTGCTCAAAAACTCTATTGGTTCAAACAAAGTTTAGGAGTGAAAGCGTCC STALLTAQKLYWFKQSL

AATGAGGAGTTTCTTTCTCCAATCAGCGAAATATTAAGCATATCTTTAAAATGGT GVKASNEEFLSPISEILSI

ATGCAACTCAAATGAAAAAATTTATGGATTTGTCATCTCCGGCTTTCTTCTCCAG SLKWYATQMKKFMDLSS

CGAAATAATATTATACTCATTACCTCCGAAAAAGCAAGCAAAGAGACTTCTTGA PAFFSSEIILYSLPPKKQA

GAATTTTCATGCTACCTTATTATCCTCTGTAACTGGTATAATATCGTTAAAAGAA KRLLENFHATLLSSVTGII

TGTCTAGACTTAGCAGAAAAGTACTACAGCGAAAGCGGCGAAAAACTCACATA SLKECLDLAEKYYSESG

TCCTGAACATTTATTATTAAACGTGTGTCTCTGCTCGGGTGCATCTGCCACTCA EKLTYPEHLLLNVCLCSG

ATCAATTATAAGAGGTGATTCAAAGTTTCTAAGGAAAGATAGATATGATC ASATQSIIRGDSKFLRKD

RYDPTSQELKKIENVALL

NAMYYYHKLSTICSGTRT

LQALLLLNRYFQLTYDTE

IYDL10 |33 GCAACATGAACAAGGCACCTTCGAGAAACATGACAGAGTAGAAGATATTTGTG 34 QHEQGTFEKHDRVEDIC 5w AAAGAATCTTTGAGCAAGGCCAAGCTCTCAAAGAAGACGAACGATATAAAGAG ERIFEQGQALKEDERYK GCTCGCGATTTATTTTTAAAGATATATTACAAGGAAGAATTTTCATCTGACGAAA EARDLFLKIYYKEEFSSD GTATAGAAAGGCTCATGACATGGAAATTTAAATCACTTATTGAGATATTACGTTT ESIERLMTWKFKSLIEILR AAGAGCCCTACAACTCTATTTTCAGAAAAACGGTGCACAGGATTTAGTTCTACA LRALQLYFQKNGAQDLV AATTTTAGAAGACACGGCAACTATGTCGGTTTTTTTACAAAGAATAGACTTTCAA LQILEDTATMSVFLQRID ATTGATGGAAATATATTTGAATTACTTTCTGATACTTTTGAGGTATTGGCACCCA FQIDGNIFELLSDTFEVLA AGTGGGAAAGAGTATTCTTGTTCGACATCGAGAAAGTTGATAGGGAAAATATGA PKWERVFLFDIEKVDRE TCTGCAAAATTGATTTCCAAAAAAACTTCATGGATCAGTTTCAATGGATTTTAAG NMICKIDFQKNFMDQFQ AAAGCCTGGCAAGGATTGTAAACTCCAAAATCTCCAGCGTATAATTAGAAAAAA WILRKPGKDCKLQNLQRI GATATTTATTGCCGTTGTTTGGTATCAAAGGTTAACCATGGGGAATGTATTCAC IRKKIFIAVVWYQRLTMG CCCGGAAATTTCTTCCCAAATAGAGATTCTTGTGAAAGATAATGAATGCTCTTC NVFTPEISSQIEILVKDNE FTTGAGGAAAATAATGATTTGGAAAGTGTATCTATGTTACTGCAGTATTACATA CSSFEENNDLESVSMLL TTGGAGTATATGAACACTGCACGAATAAACAATAGAAGGTTGTTTAAGAAGTGC QYYILEYMNTARINNRRL ATTGACTTTTTTGAAATGTTGATATCTAAGTCGCTTACTTTCTCACAAGAATCTG FKKCIDFFEMLISKSLTFS GACTGATGGTAATATTATATACCTCTAAGATTGTATTCATTTTAGACTCTGATTC QESGLMVILYTSKIVFILD AGAGAATGATTTATCCTTTGCGCTGATGAGATATTATGATCGGAAAGAAGAGCT SDSENDLSFALMRYYDR AAAAAATATGTTCCTCTATATCTTGAAACACTTGGAAGAGATGGGAAAACTTCG KEELKNMFLYILKHLEEM GGAGAGGGATATTACCTCTTTGTTTCACAAGTTCATTCTTAGCGGC GKLRERDITSLFHKFILSG

FIFTSMILEAISTDKINPFG

FEQVKIALGSPIVNV

YDL10 35 GTCAGCACCAACCCCCGCGGCTCCTGGCGCGTCCGTTCCTGCTACAGCAGCA 36 SAPTPAAPGASVPATAA 5w CCAGGAACAGAAGCAGGGATCGTTCCGGTTTCAGCAAACACTCCAAAAAGCTT PGTEAGIVPVSANTPKSL

GAATAGCAATATTAATATCAACGTAAATAATAACAATATTGGCCAACAGCAAGTT NSNININVNNNNIGQQQV

AAGAAGCCAAGAAAGCAAAGAGTGAAAAAAAAGACCAAAAAGGAATTGGAACT KKPRKQRVKKKTKKELE

AGAACGTAAAGAAAGGGAGGATTTTCAGAAACGACAACAAAAACTTTTAGAGG LERKEREDFQKRQQKLL

ATCAACAAAGGCAACAGAAATTGCTATTAGAGACAAAATTACGTCAACAATATG EDQQRQQKLLLETKLRQ

AAATCGAACTAAAAAAATTGCCTAAAGTCTACAAGAGATCAATTGTTAGGAACT QYEIELKKLPKVYKRSIV

ACAAACCCCTAATCAACCGCCTCAAGCATTACAATGGTTACGATATCAATTACA RNYKPLINRLKHYNGYDI

TCTCTAAAATAGGAGAGAAAATAGATTCCAACAAGCCAATTTTTCTCTTCGCGC NYISKIGEKIDSNKPIFLFA

CAGAGTTAGGTGCAATTAATTTACATGCTTTATCAATGTCCCTCCAATCGAAGA PELGA1NLHALSMSLQSK

ATCTTGGAGAAATAAACACCGCCTTGAACACCTTGTTGGTCACAAGCGCTGAC NLGEINTALNTLLVTSAD

TCGAACTTAAAAATATCTCTGGTCAAATACCCTGAATTATTAGACTCCTTGGCAAl SNLKISLVKYPELLDSLAI

TACTCGG L

IYGR05 |51 CAGATTCCTTTCAACTGGAGGATTTTGGCGAGGCGGTACGAATGGCACAATGT 52 RFLSTGGFWRGGTNGT

2w CTCGCACAATCAACAACGTAAATCCTTTCAAATTAAAATTCATACCGAAGACAG MSRTINNVNPFKLKFIPK

TGCCCGCAGCTGCAGACTCTGTCTCTCCAGATAGTCAACGTCCAGGTAAGAAG TVPAAADSVSPDSQRPG

CCCTTCAAATTCATAGTTTCCAATCAGAGCAAGAGTAGCAAGGCTTCTAAAAGC KKPFKFIVSNQSKSSKAS

CCGAAGTGGTCGAGCTACGCATTCCCTTCGCGTGAGACCATCAAATCTCATGA KSPKWSSYAFPSRETIKS

GGAGGCCATCAAGAAGCAGAATAAAGCTATAGACGAGCAAATAGCTGCTGCAG HEEAIKKQNKAIDEQIAA

TATCCAAGAATGACTGCTCTTGCACAGAACCTCCCAAGAAAAGAAAGAGGAAA AVSKNDCSCTEPPKKRK

TTGAGACCAAGAAAGGCGCTGATCACCCTGAGTCCGAAGGCAATCAAGCATTT RKLRPRKALITLSPKAIKH

AAGGGCACTGCTAGCTCAGCCGGAACCTAAATTGATTAGAGTTAGCGCTAGAA LRALLAQPEPKLIRVSAR

ACCGTGGATGTTCAGGACTAACGTACGATCTACAATATATCACCGAGCCGGGG NRGCSGLTYDLQYITEP

AAATTCGATGAGGTAGTAGAACAAGATGGCGTTAAAATTGTCATCGATTCAAAG GKFDEWEQDGVKIVIDS

GCGTTATTCAGCATCATTGGAAGTGAAATGGACTGGATCGACGACAAGTTGGC KALFSIIGSEMDWIDDKL

CTCTAAGTTTGTCTTCAAGAATCCAAACTCCAAGGGCACATGCGGTTGTGGCG ASKFVFKNPNSKGTCGC

AGAGTTTCATGGTTTAAAAACCTTCTGCACCATTTTTAGAAAAAAAGAATCTACC GESFMV

TATTCACTTATTTATTCATTTACTTATTTATTTACATATTTATCATACATATTAACA

TTGAACCCTCCATCGTGGTAGTGTTTGCTGTTCCTAACTTTTCTTTCGTTGTTCT

TGTAGATATATATTTTTCCAGAATTTTCTAGAAGGGTTATTAATTACAATCTTAAA

CGTTCCATAAGGGGCCGCGATTTTTTTGTTCAATTTTCAACAGGGGGCCCATCT

CAAAGAACTGCAAATTATATCACAGTAAAAGGCAAAGGGGCGCAAACTTATGC

AA

YGR05 53 GTTTGGCCCGGACGGTGGTGCATTAAACAACGACTCGAATAACCAAGACGAGC54 FGPDGGALNNDSNNQD 2w TGTTGAGGATGATGGAAAACCCCATCTTCCAATCGCAAATGAACGAGATGTTG ELLRMMENPIFQSQMNE

AGTAACCCTCAGATGTTGGACTTTATGATCCAGTCCAACCCGCAATTGCAGGC MLSNPQMLDFMIQSNPQ

CATGGGTCCACAAGCCAGGCAAATGCTACAAAGTCCCATGTTTAGACAGATGC LQAMGPQARQMLQSPM

TCACCAATCCTGATATGATTAGACAGAGCATGCAATTCGCAAGAATGATGGACC] FRQMLTNPDMIRQSMQF

CTAATGCCGGTATGGGCTCTGCAGGTGGGGCTGCCTCTGCCTTCCCCGCTCC ARMMDPNAGMGSAGGA

TGGTGGCGATGCTCCAGAGGAAGGCTCCAACACGAACACTACTTCCTCATCCA ASAFPAPGGDAPEEGSN

ACACAGGGAACAACGCAGGGACTAATGCAGGTACCAACGCAGGCGCTAACAC TNTTSSSNTGNNAGTNA

AGCTGCAAACCCATTTGCGTCTCTTCTGAACCCTGCATTAAACCCCTTTGCTAA GTNAGANTAANPFASLL

CGCGGGAAACGCTGCATCCACCGGGATGCCCGCCTTCGATCCTGCATTGCTA NPALNPFANAGNAASTG

GCGTCTATGTTCCAACCCCCTGTACAAGCAT MPAFDPALLASMFQPPV

QA

TCAACGATGGGTTTATGAAGCTCTCCGAAGCGTTAGAAAACGCTGACAAGAAG NENNTINDGFMKLSEALE

GCAAGACAAGAGATCAGGTCCAAAATGGAATTGAAGCGGCTTGCTATGGAACA NADKKARQEI RSKMELK

GGAAATGCTTGCTAAAGAATCTAAATTGAAAGAATTGAGCCAACGAGCCAGATA RLAMEQEMLAKESKLKE

CCACAACGGGACTCCGCAGACGGGAGCAATAGTTAAGCCCAAAAAGCAAACG LSQRARYHNGTPQTGAI

AGCACAGTGGCCAGACTAAAAGAGCTGGCGTACTCTCAAGGAAGAGACGTATC VKPKKQTSTVARLKELAY

CGAAAAGATAATTCTGGGCGCAGCAAAGCGTTCAGAACAACCGGATCTGCAGT SQGRDVSEKIILGAAKRS

ACGATTCAAGATTTTTCACAAGAGGGGCAAATGCCTCCG EQPDLQYDSRFFTRGAN

AS

YJR09 67 GTACAAAATTTCCAAACCCATAATACCGCAGCATATACTGACACCTAAGAAAAC 68 YKISKPIIPQHILTPKKTVK Oc GGTGAAGAACCCATATGCTTGGTCTGGTAAAAACATTTCGTTAGACCCCAAAGT NPYAWSGKNISLDPKVN

GAACGAAATGGAGGAAGAGAAAGTTGTGGATGCATTCCTGTATACTAAACCAC EMEEEKVVDAFLYTKPP

CGAATATTGTCCATATTGAATCCAGCATGCCCTCGTATAATGATTTACCTTCTCAl NIVHIESSMPSYNDLPSQ

AAAAACGGTGTCCTCAAAGAAAACTGCGTTAAAAACGAGTGAGAAATGGAGTT KTVSSKKTALKTSEKWS

ACGAATCTCCACTATCTCGATGGTTCTTGAGGGGTTCTACATACTTTAAGGATT YESPLSRWFLRGSTYFK

ATGGCTTATCAAAGACCTCTTTAAAGACCCCAACTGGGGCTCCACAACTGAAG DYGLSKTSLKTPTGAPQ

CAAATGAAAATGCTCTCCCGGATAAGTAAGGGTTACTTCAATGAGTCAGATATA LKQMKMLSRISKGYFNE

ATGCCTGACGAACGATCGCCCATCTTGGAGTATAATAACACGCCTCTGGATGC SDIMPDERSPILEYNNTP

AAATGACAGCGTGAATAACTTGGGTAATACCACGCCAGATTCACAAATCACATC LDANDSVNNLGNTTPDS

TTATCGCAACAATAACATCGATCTAATCACGGCAAGACCCCATTCAGTGATATA QITSYRNNNIDLITARPHS

CGGTACTACTGCACAACAAACTTTGGAAACCAACTTCAATGATCATCATGACTG VIYGTTAQQTLETNFNDH

CAATAAAAGCACTGAGAAACACGAGTTGATAATACCCACCCCATCAAAACCACT HDCNKSTEKHELIIPTPS

AAAGAAAAGGAAAAAAAGAAGACAAAGTAAAATGTATCAGCATTTACAACATTT KPLKKRKKRRQSKMYQH

GTCACGTTCTAAACCATTGCCGCTTACTCCAAACTCCAAATATAATGGAGAGGC LQHLSRSKPLPLTPNSKY

TAGCGTCCAATTAGGGAAGACATATACAGTTATTCAGGATTACGAGCCTAGATT NGEASVQLGKTYTVIQD

GACAGACGAAATAAGAATCTCGCTGGGTGAAAAAGTTAAAATTCTGGCCACTC YEPRLTDEIRISLGEKVKI

ATACCGATGGATGGTGTCTGGTAGAGAAGTGTAATACACGAAAGGGTACTATT LATHTDGWCLVEKCNTR

CACGTCAGTGTTGACGATAAAAGATACCTCAATGAAGATAGAGGCATTGTGCC KGTIHVSVDDKRYLNED

TGGT RGIVPGDCLQEYD

YNL26 |155 AGATTTCCTCCGGATAATGACCAAAGACCCTTTAGATGTGAAATTTGTTCACGA 156 RFPPDNDQRPFRCEICS Oc GGTTTCCACAGACTTGAACATAAAAAAAGGCACGGAAGAACGCACACTGGCGA RGFHRLEHKKRHGRTHT

GAAGCCTCACAAATGTACCGTTCAGGGCTGTCCGAAAAGCTTCAGCCGAAGCG GEKPHKCTVQGCPKSFS

ATGAACTAAAAAGACATTTGAGGACACATACTAAAGGCGTCCAAAGGCGCAGA RSDELKRHLRTHTKGVQ

ATAAAATCCAAGGGCTCGCGAAAAACCGTTGTGAATACTGCTACCGCCGCCCC RRRIKSKGSRKTWNTAT

TACCACCTTCAATGAAAACACTGGTGTTTCGCTCACGGGGATAGGTCAATCTAA AAPTTFNENTGVSLTGIG

AGTGCCACCTATTCTTATCTCCGTTGCTCAGAATTGCGATGACGTGAATATACG QSKVPPILISVAQNCDDV

AAATACTGGAAATAATAATGGCATTGTGGAGACACAGGCACCTGCAATTTTAGT NIRNTGNNNGIVETQAPA

GCCTGTGATAAATATTCCAAATGACCCTCATCCGATTCCAAGTAGCCTCTCCAC ILVPVINIPNDPHPIPSSLS

TACTTCTATCACCTCCATTGCATCAGTATATCCCTCTACTTCTCCATTCCAGTAC TTSITSIASVYPSTSPFQY

CTGAAAAGCGGGTTTCCTGAAGATCCTGCATCTACACCGTATGTACATTCGTCC LKSGFPEDPASTPYVHS

GGAAGTTCTTTAGCCCTGGGTGAATTGTCTTCAAACTCCTCTATATTTTCGAAAT SGSSLALGELSSNSSIFS

CTAGGAGGAATTTGGCCGCCATGAGTGGTCCTGATTCTTTGAGTAGTTCTAAAA KSRRNLAAMSGPDSLSS

ACCAATCCAGTGCTTCGCTTCTTTCTCAAACTTCACATCCATCAAAGAGCTTTTC SKNQSSASLLSQTSHPS

AAGACCGCCAACAGACTTAAGTCCTCTGCGAAGAATCATGCCTTCTGTAAACA KSFSRPPTDLSPLRRIMP

CAGGAGACATGGAAATTTCAAGGACAGTATCCGTTTCGAGCAGTTCATCATCA SVNTGDMEISRTVSVSS

CTCACTTCTGTTACGTATGATGACACCGCGGCTAAAGACATGGGCATGGGAAT SSSSLTSVTYDDTAAKD

ATTTTTTGATAGGCCACCTGTAACACAGAAAGCTTGCAGGAGCAATCATAAGTA MGMGIFFDRPPVTQKAC

CAAGGTTAATGCTGTTAGCAGAGGGAGACAACATGAAAGGGCACAATTTCATA RSNHKYKVNAVSRGRQ

TAT HERAQFHISGDDEDSNV

HRQESRASNTSPNVSLP

PIKSILRQIDNFNSAPSYF

SK

YNL26 157 CCAGGTCGAGTTGA1 ^"GTTCGTACCGTTCGAAGATTGAGACCGAACTAACTAA! 158 QVELICSYRSKIETELTKI Oc GATCTCCGACGATAT TGTCCGTGCTAGACTCCCACTTAATTCCATCAGCCAC SDDILSVLDSHLIPSATTG CACTGGCGAGTCCAAGGTTTTCTACTATAAGATGAAGGGTGACTACCACCGTT ESKVFYYKMKGDYHRYL ATTTGGCTGAATTTTCTAGTGGCGATGCTAGAGAAAAGGCCACAAACGCCTCTTi AEFSSGDAREKATNASL TAGAAGCATACAAGACCGCTTCTGAAATTGCCACCACAGAGTTACCCCCAACT EAYKTASEIATTELPPTH CACCCAATCC PI

YNL26 175 ffGCTCTTACGGACATCACCACAGGCAGCTCGTTAATTGATACAAAAACACCTAA 176 ALTDITTGSSLIDTKTPKF Oc GTTCGTCACAGAAGTAACACTTGAAGACGCTTTACCCAAAACATTCTATGATAT VTEVTLEDALPKTFYDMY

GTATTCTCCCGAAGTTCTGATGTCTGATCCAGCAAATATACTTTATAACGGACG SPEVLMSDPANILYNGR TCCTAAGTTTACAAAGCGCGAATTGCTGGACTGGGATCTAAACGATATACGATC PKFTKRELLDWDLNDIRS CTTGTTAATTGTGGAACAATTAAGGCCAGAATGGGGTTCCCAGTTACCGACGG LLIVEQLRPEWGSQLPTV TAGTGACCTCCGGTATAAACTTACCGCAATTCAGACTACAATTACTTCCCCTAA VTSGINLPQFRLQLLPLS GTTCCAGTGATGAGTTTATAATAGCGACATTGGTTAACTCAGACTTATACATAG SSDEFIIATLVNSDLYIEA AAGCAAATCTAGACCGCAATTTTAAGTTGACAAGCGCAAAATATACAGTTGCAT NLDRNFKLTSAKYTVASA CAGCAAGAAAAAGACATGAAGAAATGACTGGGTCAAAGGAACCCATTATGCGT RKRHEEMTGSKEPIMRL CTATCAAAGCCTGAATGGAGAAATATAATTGAGAACTATTTATTAAATGTTGCCGI SKPEWRNIIENYLLNVAV TCGAGGCCCAATGCAGATATGACTTTAAACAAAAGCGCTCCGAATACAAGAGA EAQCRYDFKQKRSEYKR TGGAAATTACTAAATTCAAATTTGAAAAGGCCTGACATGCCGCCTCCAAGCCTC WKLLNSNLKRPDMPPPS ATACCGCATGGTTTTAAAATACATGACTGCACTAACTCTGGTAGTCTTTTAAAAA LIPHGFKIHDCTNSGSLL AGGCTTTAATGAAAAATTTGCAACTAAAAAATTATAAAAATGATGCTAAGACATT KKALMKNLQLKNYKNDA AGGTGCTGGTACACAGAAAAATGTCGTTAATAAGGTTTCTCTAACTTCAGAGGA KTLGAGTQKNWNKVSL GAGGGCTGCCATCTGGTTTCAATGCCAAACACAGGTTTATCAAAGGTTGGGGT TSEERAAIWFQCQTQVY TAGATTGGAAGCCTGATGGAATGTCCTAGTAAATTTTTACAACAACGCAAAAAC QRLGLDWKPDGMS GAAATACCCCACTATTTAAC

YNL26 177 CCGCCAAGCGATAATAGTAATAGTACGGCCGGTGGAGCCAATGGAAGTAACTC 178 PPSDNSNSTAGGANGS Oc AGGAACACCAACTTCTACTAGCGGTAAGAAGAGAAACAAGCTTATAAAATCTTG NSGTPTSTSGKKRNKLIK

TGGCTTTTGCAGAAGGAGAAAACTTCGTTGTGATCAGCAAAAACCTATGTGTTC SCGFCRRRKLRCDQQK

TACATGCATTTCTAGAAACTTAACAACCTGTCAATATGCTGAAGAATTCAACAAA PMCSTCISRNLTTCQYA

AACATCGAAAAGAAAGCTACCTATGGTCCCTATCCTAACGCCGATTTACTTAAG EEFNKNIEKKATYGPYPN

AAAGTTG G CTAGAAMCAAAATACGTATTCTAG GCTGAAAAAAACACT ADLLKKVEELENKIRILEA

AATTCGTCTGCGAGCTCCATGTACACTTCGCCAAATTTCCCTCCTTTAGGCACT EKNTNSSASSMYTSPNF

AGTGTAGGTAGAGGTTCTACTGAAACTTCATCTCCATTACCCGATGGTGTAATA PPLGTSVGRGSTETSSP

AATCCATATGCCGACCGGTACTACCTACAAAGTAAACATTCCGGAAGATCAACA LPDGVI NPYADRYYLQSK

CTATACGGCCCCACTTCTATGAGAACCCAAATTGCAAATAGTAATTGGGGGTTC HSGRSTLYGPTSMRTQI

ATTGAAAAATATAAACAATTATGGGCGAAGGTTAAAGTAGAAAGAAATAAATGG ANSNWGFIEKYKQLWAK

AAGCAAAACAACCAAAAAACGATGTGCAGGGAACTAGGCCTTTTGGATGAGTC VKVERNKWKQNNQKTM

GGATTGGCAGCCAGATCCATTAATCAAACAGATATGTCGTTTCCTACCATCATA CRELGLLDESDWQPDPL

TAACAAAGCTTTGTCTATTTTAGATGATTTCTTTAATGATGGAGCATGCAATGAG IKQICRFLPSYNKALSILD

ATC DFFNDGACNEI

YER05 [231 GCCGACCAAGTCAACTTTTAGTCGCTGGAAGAAGGCGGACCTAATTGACCTGG 232 PTKSTFSRWKKADLIDLA

CCAACAAGCTGGAAATTGACGGCTTCCCCAATTATGCCAAGAAGAGTGATATG NKLEIDGFPNYAKKSDMI

ATCGATTACCTCGAATCGCACTTGAATCATCTTGAGAAACCTGTGGATTTTAAA DYLESHLNHLEKPVDFK

GACGACTACCCGGAACTAAGGTCCTTTTATGAGTCGATGACAGTGGACCAGTC DDYPELRSFYESMTVDQ

AAAGGATGAACGGAACGAATACGGATCCGGATCTGGAAACGGGTCTGGGTCT SKDERNEYGSGSGNGS

GGATCCTGTGACACCGCCACAAATGATTCCGATCTGGAGAAAGCGTACATCAA GSGSCDTATNDSDLEKA

GGAGGATGATGATGAGAAACCGCAATCAGGCGATGAAACAAGCGCGACAAAG YIKEDDDEKPQSGDETS

CCGCTTTCCAGCAGGAATGCAAATTCAAACGCCAAAACGAACTTCAATTTGTTG ATKPLSSRNANSNAKTN

GATTTTAGCACAGATAACGACTCGTCCACCTCTGCGTTTACTAAGTTCAAGTTT FNLLDFSTDNDSSTSAFT

AACTTTCAGGAATACCTCTCCGATATTAGATACCAAACGCAGAAATTGAATGAA KFKFNFQEYLSDIRYQTQ

AATGTTCAGGACTATCTTTCCACTATTTCCGCCGTCGATACGATTTTTTCCTTGT KLNENVQDYLSTISAVDT

TGGAATTCTCTTTTCTGGTGAGAAACATATTAGCCGCCGGGCAGCCAACTTCTT IFSLLEFSFLVRNILAAGQ

CTTCTTCGCTCGCATCGTCGCTGGAAGCCGCCGTTGCCGCCCATAATAAATAT PTSSSSLASSLEAAVAAH

CAATACACGCTCGATTTCTGTTTACCAATACTCACGTGGCTGCTCTTCTTTAGG NKYQYTLDFCLPILTWLL

GG FFR

YER05 233 GAGATCGATAACAAGGTATTCGACCTGGAAATTTTAGATACAGCGGGTATAGC 234 EIDNKVFDLEILDTAGIAQ 7c ACAATTTACTGCAATGAGAGAATTATACATAAAGTCAGGAATGGGGTTCCTGTT FTAMRELYIKSGMGFLLV

GGTATATTCAGTAACAGATCGGCAATCTTTGGAAGAATTAATGGAGCTAAGAGA YSVTDRQSLEELMELRE

ACAGGTCCTTAGGATCAAAGATTCCGATCGCGTTCCAATGGTTCTAATAGGTAA QVLRIKDSDRVPMVLIGN

CAAGGCTGATCTAATCAATGAAAGGGTAATAAGTGTGGAAGAAGGTATAGAGG KADLINERVISVEEGIEVS

TAAGCAGTAAATGGGGTAGAGTTCCCTTTTATGAAACAAGCGCTTTGTTGAGGAl S KWG RVPF YETS ALL RS

GCAATGTGGATGAAGTGTTCGTTGATTTAGTTAGGCAAATCATTCGGAATGAAA NVDEVFVDLVRQIIRNEM

TGGAAAGTGTTGCAGTTAAAGACGCAAGAAATCAAAGTCAACAATTTAGCAAAA ESVAVKDARNQSQQFSK

TCGAGTCTCCATCAACCAGGCTCCCTAGTTCTGCGAAACAGGATACGAAACAA IESPSTRLPSSAKQDTKQ

TCAAACAATAAGCAATCATCAAAAGGTTTATATAACAAATCTTCACAAGGACAAG SNNKQSSKGLYNKSSQG

CTAAAGTTAAACAATCTACTCCGGTTAATGAAAAGCACAAACCGTCACATGCCG QAKVKQSTPVNEKHKPS

TTCCGAAATCTGGTTCTAGCAACAGGACAGGAATTAGCGCTACTTCACAACAAA HAVPKSGSSNRTGISAT

AGAAAAAGAAGAAAAACGCTTCCACTTGCACTATTCTATAGTCACTTAATTTTAT SQQKKKKKNASTCTIL

TATAAATGAATCAAGATATCAGATAAAAAGACTTTACTTGAAATAGTTTTATTATAl

GTTCTAAAAGGTTTAGTTTAAAGTATTAGCATACGTTGTATAAGTTTTTAAAGAA

ATCAATTAATAATGTTTGAAAATAAATTTAAACCCAAAAAAAATGAAATGTTAAAA

ATATGGACGCAACCGGAATCGAACCGATGACCTCTTCCTTGCAAGGGAAGCGC

GCTACCAACTGCGCCATGTGCCCGCAATCTATGGGATTTTACGGTAGATGCTG

CGTTACGTATAAAAAATATTAAATGAACAGATCCACACTCTCGTAACTAA

YKL 9 I275 IGCGCCAAGACCACATGCTTGTCCTATCTGTCATAGAGCTTTTCACAGACTGGA |276 APRPHACPICHRAFHRL 5w ACATCAGACGAGACACATGAGAATTCATACAGGTGAGAAGCCTCACGCGTGTG EHQTRHMRIHTGEKPHA

ACTTCCCCGGATGTGTGAAAAGGTTCAGTAGAAGCGATGAACTGACGAGACAC CDFPGCVKRFSRSDELT AGAAGAATTCATACAAACTCCCACCCTCGAGGTAAAAGAGGCAGAAAGAAGAA RHRRIHTNSHPRGKRGR GGTTGTGGGCTCTCCAATAAATAGTGCTAGTTCTAGTGCTACCAGTATACCAGAl KKKWGSPINSASSSATS TTTAAATACGGCAAATTTTTCACCGCCATTACCACAGCAACACCTATCGCCTTT IPDLNTANFSPPLPQQHL AATTCCTATTGCTATTGCTCCGAAAGAAAATTCAAGTCGATCTTCTACAAGAAAA! SPLIPIAIAPKENSSRSST GGTAGAAAAACCAAATTCGAAATCGGCGAAAGTGGTGGGAATGACCCATATAT RKGRKTKFEIGESGGND GGTTTCTTCTCCCAAAACGATGGCTAAGATTCCCGTCTCGGTGAAGCCTCCAC PYMVSSPKTMAKIPVSV CTTCTTTAGCACTGAATAATATGAACTACCAAACTTCATCCGCTTCCACTGCTTT KPPPSLALNNMNYQTSS GTCTTCGTTGAGCAATAGCCATAGTGGCAGTAGACTGAAACTGAACGCGTTAT ASTALSSLSNSHSGSRL CGTCCCTACAAATGATGACGCCCATTGCTAGCAGTGCGCCAAGGACTG KLNALSSLQMMTPIASSA

PRT

YKL19 277 GACTCCAGCAACTTTTCTACAGGTTTCAGCGGCAAGATTCGTAAGCCAAGGTC 278 DSSNFSTGFSGKIRKPR 5w GAAAGTAAGTAAAGCGTGCGATAACTGTAGAAAAAGAAAGATAAAATGTAATGG SKVSKACDNCRKRKIKC

GAAGTTTCCCTGCGCAAGCTGTGAGATATATTCATGTGAGTGCACGTTCAGCA NGKFPCASCEIYSCECTF

CTAGACAAGGTGGCGCTCGAATAAAAAACCTTCACAAGACGAGTTTGGAAGGT STRQGGARIKNLHKTSLE

ACAACCGTACAAGTCAAAGAGGAAACAGATTCCAGTTCGACTTCTTTTTCTAAT GTTVQVKEETDSSSTSF

CCTCAGCGATGTACAGACGGGCCATGCGCAGTGGAACAACCAACGAAATTTTT SNPQRCTDGPCAVEQPT

TGAGAATTTCAAGCTAGGTGGTCGTAGTAGTGGTGATAATAGCGGAAGTGATG KFFENFKLGGRSSGDNS

GGAAGAATGACGACGATGTGAATAGAAACGGTTTTTATGAAGACGATAGCGAA GSDGKNDDDVNRNGFY

TCACAGGCAACTTTGACATCTCTACAAACCACTCTGAAAAATTTGAAGGAGATG EDDSESQATLTSLQTTLK

GCTCATTTAGGTACACATGTAACCTCAGCCATCGAGAGCATTGAACTTCAGATA NLKEMAHLGTHVTSAIES

AGTGACTTGCTTAAGCGATGGGAACCCAAAGTGAGGACCAAAGAATTAGCAA IELQISDLLKRWEPKVRT

KELA

YKL19 (279 AATGCTGCTGGTGCCAAGGAAGTTTTGAAGGAATCTGCAAAGACTATTGTCGA 280 NAAGAKEVLKESAKTIVD 5w TTCTGGCAAACTACCATCCAGCTTGTTGTCCTACTTCGTGTGAATACCGTAAGA SGKLPSSLLSYFV

AATGGAATAGAATATATACGAATGTATACGAATATTATAGAGAACGTTCTCTTTT

ATTTCTATAATGAATAGGTTCGGGTAACGGTTCCCTTTTTAGGTATTTCTAGAAGI

ATGAGAGAAGAGGGAATAATGAGAAAGGCGAAAAATAAAGGACACCTTTAACG

AAAGATCAAAGGTGTCCTTATTTACTTACAATAGCTGCAATTAGTACGACTCAAAl

AAAAGTGAAAACAAAACTGAAAGGATAGATCAATGTCTTACAGAGGACCTATTG

GAAATTTTGGCGGTATGCCAATGTCATCATCGCAAGGACCATACTCTGGCGGT

GCACAATTCAGATCAAACCAGAACCAATCCACTTCTGGCATCTTAAAGCAATGG

AAGCATTCTTTTGAAAAGTTTGCCTCCAGAATTGAGGGGCTCACTGACAATGCA

GTTGTTTATAAATTGAAGCCTTACATTCCAAGTTTGTCAAGATTTTTCATTGTGG

CCACCTTTTATGAAGATTCGTTTAGGATCTTATCACAATGGTCAGATCAAATTTT

TTATCTGAATAAGTGGAAGCATTACCCATACTTCTTTGTCGTTGTGTTTCTAGTG

GTTGTTACCGTTTCCATGTTGATTGGCGCCAGTTTGTTAGTTTTAAGAAAGCAA

ACCAATTATGCCACCGGTGTGTTATGTGCTTGCGTTATTTCTCAAGCATTAGTT

TATGGGTTGTTTACGGGTTCATCATTTGTCCTAAGAAACTTTAGTGTTATTGGTG

GGTTGTTAATTGCATTCAGCGATTCAATTGTTCAAAACAAGACAA

YOR05 281 CAGGCTGCACCACCAGCTTTGCAGCCCACCGATTTCCAGCAATCTCACATTGC 282 QAAPPALQPTDFQQSHI

7w AGAAGCCTCCAAATCACTGGTAGACTGCACAAAGCAAGCCTTGATGGAAATGG AEASKSLVDCTKQALME

CCGACACTCTCACCGACAGCAAGACAGCAAAGAAACAACAACCTACGGGAGAT MADTLTDSKTAKKQQPT

AGCACTCCCTCAGGCACGGCAACTAACAGTGCAGTTTCTACACCATTGACTCC GDSTPSGTATNSAVSTP

CAAGATAGAGCTGTTTGCTAATGGCAAGGACGAAGCCAACCAGGCGCTCTTAC LTPKIELFANGKDEANQA

AACACAAGAAACTGTCTCAGTACAGCATCGACGAAGATGACGACATTGAAAAC LLQHKKLSQYSIDEDDDI

AGAATGGTCATGCCCAAGGACTCGAAATACGACGACCAATTATGGCACGCGCT ENRMVMPKDSKYDDQL

AGATTTGTCCAACTTGCAAATCTTCAATATCAGCGCCAACATCTTCAAGTACGA WHALDLSNLQIFNISANIF

TTTTCTAACGAGACTATATTTGAATGGCAATAGCCTCACGGAACTGCCAGCGGA KYDFLTRLYLNGNSLTEL

GATCAAGAACCTAAGCAACCTACGCGTTTTGGACCTGTCGCATAATAGGTTAAC PAEIKNLSNLRVLDLSHN

ATC RLT

AATGTTGTTAATTAAGGGTAAAATATGATGATGTATTTATGATAAGTAAAGAGTA

GGGGTCACCTAACATTTTTCACAATTGCATTTACTCTTTTCACAAGTGCACGTG

GTTTCGCATTTGCATTTTTCACCACAACTCTTACATTGAGGGCTTCCG

YOR05 303 TGCTCTTACGGACATCACCACAGGCAGCTCGTTAATTGATACAAAAACACCTAA 304 ALTDITTGSSLIDTKTPKF 7w GTTCGTCACAGAAGTAACACTTGAAGACGCTTTACCCAAAACATTCTATGATAT VTEVTLEDALPKTFYDMY

GTATTCTCCCGAAGTTCTGATGTCTGATCCAGCAAATATACTTTATAACGGACG SPEVLMSDPANILYNGR

TCCTAAGTTTACAAAGCGCGAATTGCTGGACTGGGATCTAAACGATATACGATC PKFTKRELLDWDLNDIRS

CTTGTTAATTGTGGAACAATTAAGGCCAGAATGGGGTTCCCAGTTACCGACGG LLIVEQLRPEWGSQLPTV

TAGTGACCTCCGGTATAAACTTACCGCAATTCAGACTACAATTACTTCCCCTAA VTSGINLPQFRLQLLPLS

GTTCCAGTGATGAGTTTATAATAGCGACATTGGTTAACTCAGACTTATACATAG SSDEFIIATLVNSDLYIEA

AAGCAAATCTAGACCGCAATTTTAAGTTGACAAGCGCAAAATATACAGTTGCAT NLDRNFKLTSAKYTVASA

CAGCAAGAAAAAGACATGAAGAAATGACTGGGTCAAAGGAACCCATTATGCGT RKRHEEMTGSKEPIMRL

CTATCAAAGCCTGAATGGAGAAATATAATTGAGAACTATTTATTAAATGTTGCC SKPEWRNIIENYLLNVAV

TCGAGGCCCAATGCAGATATGACTTTAAACAAAAGCGCTCCGAATACAAGAGA EAQCRYDFKQKRSEYKR

TGGAAATTACTAAATTCAAATTTGAAAAGGCCTGACATGCCGCCTCCAAGCCTC WKLLNSNLKRPDMPPPS

ATACCGCATGGTTTTAAAATACATGACTGCACTAACTCTGGTAGTCTTTTAAAAA LIPHGFKIHDCTNSGSLL

AGGCTTTAATGAAAAATTTGCAACTAAAAAATTATAAAAATGATGCTAAGACATT KKALMKNLQLKNYKNDA

AGGTGCTGGTACACAGAAAAATGTCGTTAATAAGGTTTCTCTAACTTCAGAGGA KTLGAGTQKNWNKVSL

GAGGGCTGCCATCTGGTTTCAATGCCAAACACAGGTTTATCAAAGGTTGGGGT TSEERAAIWFQCQTQVY

TAGATTGGAAGCCTGATGGAATGTCCTAGTAAATTTTTACAACAACGCAAAAAC QRLGLDWKPDGMS

GAAATA

YOR05 |305 GTGGCCAAACAAGAATGAAAAAAACCACACAGTTAAAAGAGCGTTATCAACGG 306 WPNKNEKNHTVKRALST 7w ATATGACCAGCAATATTTTGAGTAGCACAAACGCGAGCTCAAACGAAGAAAATT DMTSNILSSTNASSNEEN

CTAGGAGTTCTTCTGCAGCCAATGTACGCTCCGGAACAGGTGCAAATACACTT SRSSSAANVRSGTGANT

ACTAATG G CGGCAGTACTAGAAAG AG ACTTG CGTGCACTAATTGTAGAAATAG LTNGGSTRKRLACTNCR

AAGGAAAAAATGTGATTTAGGATTTCCCTGTGGTAACTGTTCCAGGTTGGAATT NRRKKCDLGFPCGNCS

GGTTTGTAATGTTAATGACGAGGACTTAAGAAAGAAGCGGTACACTAATAAATA RLELVCNVNDEDLRKKR

TGTCAAGTCTTTGGAGAGCCATATTGCTCAACTGGAGACCAACTTAAAAAACCT YTNKYVKSLESHIAQLET

AGTTCAGAAGATCTACCCTGACGATGAGCAAATACTGAACCGAATGATGGTAG NLKNLVQKIYPDDEQILN

GTGATGTATTATCAGCTCTACCAGACAGTTCACAAGTCTCAATCAATTATACTG RMMVGDVLSALPDSSQV

ACCAAACTCCCTCTCTTCCAATTCCCGCAACCAGAGGTACATTCATTATTGAAA SINYTDQTPSLPIPATRG

ACGATAAGGTCAGTCAACCTCTATCGTCCTTTAACCAACAAACAGAGCCATCTA TFIIENDKVSQPLSSFNQ

CTCTAAACTCGGGTATCTTCAACACCCAAAAACAAAATTTCGAAGAATCCCTTG QTEPSTLNSGIFNTQKQ

ATGATCAGTTACTTTTACGAAGATCGTTAACACCGCAAGGTGAAAAAAAGAAGA NFEESLDDQLLLRRSLTP

AACCGTTGGTAAAAGGTAGTCTTTATCCTGAAGGACCTGTCAGTTACAAACGGAl QGEKKKKPLVKGSLYPE

AGCACCCCGTAAAATCGGACAGTTTATTGCCTGTGTCTTCGTTAACAGCTGCTA GPVSYKRKHPVKSDSLL

CAGACCCATCTACTTTTTCTGACGGTATAACTGCTGGTAATTCCGTCCTAGTTA PVSSLTAATDPSTFSDGI

ATGGTGAACTGAAAAAACGTATATCCGACTTGAAGACCACCGTAATAGTAAGAG TAGNSVLVNGELKKRISD

GACTAAACGATGATAATCCCAACTCTATCAATAACGATCCCAGGATTTTAAAAT LKTTVIVRGLNDDNPNSI

CTCTTTCCAATTTCTATAAGTGGCTGTATCCAGGTTATTTTATTTTTGTTCACAG NNDPRILKSLSNFYKWLY

A PGYFIFVHRESFLYGFFN

HSKNNYEDSSYCSVELIY

AMCAVGSRLTPDLQEYS

EVYYQRSKKTLLQLVFD

EQSTARITTVQALFCLAF

YELGKGNNQLGWYFSG

LAIRVGYDMGFQLDPKV

WYVDDNNLQLTQSELEI

RSRIYWGCYIADHFICLM

LGRTSTLSVSNSTMPES

DELPEVNGTEEFRFIGRH

VLQISLPLKNLIILSRLVQI

FTSKIFIESEDIARKLKYL

NTFNSQVYNWRQSLPEF

LQWSKTLIENDDVSTDPT

YPL04 1313 GAGGACGCAACTGCGCCTGACTTTGCTTTGCTCTCCGGGTTCAGCATCGAATG 314 RTQLRLTLLCSPGSASN 8w AAAGCTCTGTCTGCTCTTCTAACGCCAGCGACTTAGACATGTCCCTTTTATCTA ESSVCSSNASDLDMSLL

CTCCTTCAAGTCTCTTCCAGATGGCAGGTGAGACCAAAAGCAATCCTATAATTA STPSSLFQMAGETKSNPI

TACCCGACAGTCAAGACGATAGTATACTTAGTAGCGACCCCTTTTAAGTAGGTA IIPDSQDDSILSSDPF

ACCCCCCCTGGATTGCATAGCCATTGCATAGCCTTAGACATCAAACTTTATTTA

ATCACATTATCCTTCTATGTATCTTTTTCCCCCGCTCGATTTCTTCTAGAACATT

ACGGAAAATAAAGGAAAAAAATGACTGGAGCATCGAATCTGTAGACTAAAAAG

GTAATGACGCGTTCTTCGTTCCCACAAGTATGTGAAATCGTTTGAACGCTTTTA

TATACTGATAGGAATAGATTATAATAGTATTCAACATTCATCAAACAGTTTATAT

CGGTAATAAACCAACCTCACGATACAGTGATTATTTCTCTAAACAACACGAGCC

GTACTGTACCTACGATAAAGTAAAACACATTTTTCTTTTGCTACCAGTGG

YDR24 315 GCCAGCACCACGAATTCCGAGTAAAGACGCCATTATATCAAGGTTGGAAAAAG 316 PAPRIPSKDAIISRLEKDM 7w ATATGTTTTATTGGAAAGATAAAGCTATGAAGCTACTAACAGAGAGAGAGGTGA FYWKDKAMKLLTEREVN

ATGAATCAGGCAAGAGATCAGCAAGTCCGATCAATACAAACAATGCTAGCGGG ESGKRSASPI NTN NASG

GACAGTCCTGATACCAAGAAGCAGCATAAAATGGAACCTATATATGAACAAAGT DSPDTKKQHKMEPIYEQ

GGTAACGGGGATATAAACAATGGTACCAGAAATGATATTGAAATCAACTTGTAT SGNGDINNGTRNDIEINL

AGAAGTCATCCAACCATGATCATGAGTAAAGTCATGAAAAGAGAAGTTAAGCC YRSHPTMIMSKVMKREV

GTTATCTGAAAATTATATTATAATTCAGGACTGTTTTCTAAAAATCCTGGTCACT KPLSENYIIIQDCFLKILVT

TCAGTGTTCCTTGACACTTCAAAGAACACGATGATACCGGCATTGACGGCAAA SVFLDTSKNTM I PALTAN

CGCGAATATTACAAGAGCCCAGCCTAGCGTA ANITRAQPSV

YDR24 317 AAATGGAGGATATACCAAACCACAAAAATATGTGCCAGGGCCAGGTGATCCTG 318 NGGYTKPQKYVPGPGD 7w AACTTCCACCCCAACTATCCGAATTTAAAGATAAAACATCGGATGAAATCTTGA PELPPQLSEFKDKTSDEI

AAGAAATGAACAGAATGCCTTTTTTCATGACCAAGTTGGATGAAACAGACGGTG LKEMNRMPFFMTKLDET

CAGGTGGTGAAAACGTGGAGTTAGAAGCTTTAAAGGCATTAGCTTATGAAGGC DGAGGENVELEALKALA

GAACCACACGAAATCGCTGAAAATTTCAAGAAGCAAGGTAACGAACTATACAAA YEGEPHEIAENFKKQGN

GCAAAAAGATTCAAGGATGCAAGGGAACTTTACTCAAAGGGCTTGGCTGTAGA ELYKAKRFKDARELYSK

ATGCGAAGATAAATCAATAAATGAGTCACTATATGCCAATAGAGCGGCATGTGA GLAVECEDKSINESLYAN

GTTAGAGCTGAAAAATTACAGGAGGTGTATCGAGGACTGCAGTAAAGCTCTAA RAACELELKNYRRCIEDC

CTATTAACCCCAAGAATGTTAAGTGCTACTATCGTACA SKALTINPKNVKCYYRT

YDR24 325 CGGCTTTACTAAGTCTATCGAGAATGAGATCTATCAAATTTTAAAAAATCTGCGT 326 GFTKSIENEIYQILKNLRY 7w TATCCGTTTTTAGAGTCAATAAATAAATCACAAATTTCGGCTGTAGGTGGCTCT/> PFLESINKSQ1SAVGGSN

ATTGGCACAAATTTCTTGGCATGTTGCATTGGATGGTACGAACAAATATTAAAC WHKFLGMLHWMVRTNIKl

TGGATATGTGCTTGAATAAAGTAGATCGTTCATTAATTAATCAAAATACACAAGA LDMCLNKVDRSLINQNT

AATAACAATTCTGAGCCAGCCTTTAAAGACTTTGGACGAACAGGACCAAAGACA QEITILSQPLKTLDEQDQ

AGAAAGATATGAGCTAATGGTGGAGAAACTGTTAATTGATTATTTTACAGAGTC RQERYELMVEKLLIDYFT

TTACAAAAGCTTTTTGAAACTTGAGGATAATTATGAGCCTTCGATGCAAGAACT ESYKSFLKLEDNYEPSM

AAAGTTAGGTTTTGAAAAATTCGTTCACATAATTAATACTGATATAGCTAATCTA QELKLGFEKFVHIINTDIA

CAAACCCAAAATGACAATCTTTATGAGAAATATCAAGAAGTAATGAAAATAAGC NLQTQNDNLYEKYQEVM

CAAAAGATCAAAACCACCAGGGAAAAATGGAAGGCTTTGAAAAGCGATTCTAAT KISQKIKTTREKWKALKS

AAGTATGAAAATTATGTCAACGCGATGAAGCAAAAGAGTCAAGAATGGCCAGG DSNKYENYVNAMKQKS

TAAACTGGAAAAGATGAAATCCGAGTGTGAACTGAAAGAAGAAGAAATTAAAGC QEWPGKLEKMKSECELK

CTTACAAAGTAATATTTCCGAATTGCACAAAATTTTAAGAAAAAAGGGAATTTCA EEEIKALQSNISELHKILR

ACTGAGCAGTTCGAATTACAAAACCAAGAAAGAGAAAAGCTGACTAGGGAACT KKGISTEQFELQNQERE

TGATAAAATAAATATCCAGTCTGATAAATTGACAAGCTCAATTAAATCCAGAAAG KLTRELDKINIQSDKLTSS

CTGGAAGCCGAGGGAATATTCAAAAGCTTACTGGATACGTTGAGGCAATACGA IKSRKLEAEGIFKSLLDTL

TTCGTCGATACAAAATTTAACCAGATCGCGTAGTCAATTGGGTCATAATGTTAA RQYDSSIQNLTRSRSQL

TGATTCATCTCTAAAAATTAACATTTCAGAGAACTTATTAGACAGAGATTTTCAT GHNVNDSSLKINISENLL

GAAGGTATCTCCTACGAGC DRDFHEGISYE

YDR24 327 GTGATCGCACCGCTGACAGATGATTATGATGTATGGACGAGAAGCAATAATTT 328 VIAPLTDDYDVWTRSNN 7w CATTGATATTAAGTTGCCTAAAGAAATAGGTGAGCAGATAAATGATGGGCAGGT FIDIKLPKEIGEQINDGQVI

GATCATAGATAACATGAATGAATTGATACAAAATACATTGCCTACAAGTCAGAT IDNMNELIQNTLPTSQMM

GATGGCACGTGAGCAGGCTGTTTTTGAAAATGATTACGATTTCTTCTTTAACGA AREQAVFENDYDFFFNE

ATACAGGGACCTGGATACTATTTATATGTGGTTGGATCTTCTAGAAAGAAGCTT YRDLDTIYMWLDLLERSF

TCCCAGTTTGGTTGCAGTGGAACACTTGGGCAGGACATTTGAAGGCAGAGAGQ PSLVAVEHLGRTFEGRE

TAAAAGCTCTGCATATATCCGGAAACAAGCCAGAATCAAACCCGGAAAAGAAG LKALHISGNKPESNPEKK

ACTATTGTTATCACAGGTGGTATACATGCCAGAGAGTGGATCAGTGTCAGTAC TIVITGGIHAREWISVSTV

CGTCTGTTGGGCACTTTATCAGTTGTTAAATCGATATGGTTCGTCTAAGAAGGA CWALYQLLNRYGSSKKE

AACCAAATACCTGGACGATTTGGACTTTTTAGTCATACCTGTGTTCAATCCAGA TKYLDDLDFLVIPVFNPD

CGGATACGCATATACGTGGTCGCATGACAGACTATGGCGTAAGAATCGTCAAA GYAYTWSHDRLWRKNR

GGACTCATGTTCCTCAATGTCTTGGTATCGACATTGATCATTCCTTTGGTTTCC QRTHVPQCLGIDIDHSFG

AGTGGGAAAAGGCACACACCCACGCTTGCAGCGAAGAGTATAGTGGGGAAAC FQWEKAHTHACSEEYS

GCCCTTCGAAGCTTGGGAAGCATCTGCTTGGTACAAGTATATTAATGAAACCAA GETPFEAWEASAWYKYI

GGGCGATTACAAAATTTATGGCTACATTGACATGCATTCGTATTCA NETKGDYKIYGYIDMHSY

YKR08 1347 GGTTTGACTAAGGGTGCCTCTGCTGGTGAAGGTTTGGGTAATGCTTTCTTGTC 348 GLTKGASAGEGLGNAFL 1c TCACATCAGATCAGTCGATTCTATCTACCAAGTCGTTCGTTGTTTCGATGATGC SHIRSVDSIYQWRCFDD

TGAAATTATCCACGTTGAGGGTGACGTTGATCCAGTTCGTGATTTAGAAATTAT AEIIHVEGDVDPVRDLEII

TAACCAAGAACTAAGATTGAAAGATATTGAATTCGCACAAAAGGCTTTGGAAGG NQELRLKDIEFAQKALEG

TGCTGAAAAGATTGCCAAAAGAGGTGGTCAATCTTTGGAAGTCAAACAAAAGAAl AEKIAKRGGQSLEVKQK

GGAAGAAATGGATTTGATTACGAAAATCATTAAATTGCTAGAGAGTGGTCAAAG KEEMDLITKIIKLLESGQR

AGTTGCTAATCACTCCTGGACTTCAAAAGAAGTTGAAATTATCAACTCCATGTT VANHSWTSKEVEIINSMF

CTTGTTGACTGCTAAGCCATGTATCTATTTGATTAATTTATCTGAAAGAGATTAC LLTAKPCIYLINLSERDYI

ATCAGAAAGAAAAACAAGCATCTGCTAAGAATCAAGGAATGGGTAGACAAGTA RKKNKHLLRIKEWVDKY

CTCTCCAGGTGACTTGATCATTCCATTCAGTGTTTCTCTAGAAGAAAGACTATC SPGDLIIPFSVSLEERLSH

TCATATGTCCCCAGAAGATGCTGAAGAAGAATTGAAGAAACTGCAGACAATATC MSPEDAEEELKKLQTISA

TG CCTTG CCAAAG ATTATCACTACC ATG AG ACAAAAGTTAG ATTTG ATTTCCTTT LPKIITTMRQKLDLISFFT

TTCACCTGCGGTCCAGATGAAGTTCGTGAATGGACCATCAGAAGAGGTACTAA CGPDEVREWTIRRGTKA

AGCTCCACAAGCTGCTGGTGTTATTCATAACGATTTAATGAATACCTTTATTTTG PQAAGVIHNDLMNTFILA

GCTCAAGTTATGAAATGTGAAGATGTCTTCGAATATAAGGACGATTCTGCCATC QVMKCEDVFEYKDDSAI

AAGGCCGCTGGTAAGTT KAAGK

YKR08 349 AGTGAGCCTGGATGAAGCATTACCCAAAACGTTTTATGACATGTATTCGCCAGA 350 VSLDEALPKTFYDMYSP 1c TATTCTATTAGCAGACCCATCCAACATTCTCTGTAACGGGCGTCCCAAGTTTAC DILLADPSNILCNGRPKF

CAAGAGAGAGTTATTGGATTGGGATTTAAACGATATAAGATCGTTATTGATAGT TKRELLDWDLNDIRSLLI

CGAGAAGTTAAGGCCCGAATGGGGTAATCAACTACCGGAAGTAATAACGGTGG VEKLRPEWGNQLPEVIT

GTGATAATATGCCCCAGTTTAGGTTACAATTATTACCACTATATTCTAGCGATGA VGDNMPQFRLQLLPLYS

GACCATAATCGCAACGTTAGTCCATTCGGATCTGTACATGGAGGCTAACTTAGA SDETIIATLVHSDLYMEA

TTATGAATTCAAACTAACCAGCGCCAAATATACAGTAGCGACCGCTAGAAAAAG NLDYEFKLTSAKYTVATA

ACATGAGCATATAACTGGTAGAAATGAAGCCGTCATGAATTTGTCGAAACCGG RKRHEHITGRNEAVMNL

AATGGAGAAATATCATCGAAAATTACCTCTTAAATATAGCAGTAGAGGCACAAT SKPEWRNIIENYLLNIAVE

GCAGGTTTGATTTCAAACAAAGATGCTCCGAATATAAGAAATGGAAGTTACAAC AQCRFDFKQRCSEYKK

AGTCCAACTTAAAAAGACCGGACATGCCCCCACCAAGCATAATACCGCGGAAA WKLQQSNLKRPDMPPP

AACAGCACAGAAACAAAATCGCTTCTGAAAAAGGCTTTATTGAAGAACATTCAG S 11 PRKNSTETKS LLKKAL

TTGAAAAACCCCAATAATAACCTTGATGAATTGATGATGAGATCAAGCGCCGCA LKNIQLKNPNNNLDELM

ACAAATCAACAGGGAAAAAACAAAGTCAGCTTATCTAAAGAAGAAAAGGCTACG MRSSAATNQQGKNKVSL

ATATGGTCGCAATGTCAGGCACAAGTTTACCAAAGATTAGGATTGGATTGGCA SKEEKATIWSQCQAQVY

GCCGGATTCAGTATCCTGAAGATACTAAAAA QRLGLDWQPDSVS

YKR08 1363 CTGAGAAGATTGAAGGAAGACGTTCTATCTGATTTACCACCTAAAATTATTCAG 364 LRRLKEDVLSDLPPKIIQD 1c GATT CTACTGTGAATTAGGTGATTTACAAAAACAACTATACATGGATTTTACTA YYCELGDLQKQLYMDFT

AAAAACAAAAAAATGTAGTTGAGAAAGATATTGAAAATTCTGAGATTGCTGATG KKQKNWEKDIENSEIAD

GCAAGCAACATA fTCCAAGCTTTACAATACATGAGAAAATTGTGTAATCATCC GKQHIFQALQYMRKLCN

AGCT ΓGGTCCTTTCACCAAATCACCCGCAATTAGCGCAAGTACAAGACTATTT HPALVLSPNHPQLAQVQ

AAAGCAAACTGGTCTAGATTTACACGATATTATCAACGCTCCAAAACTGAGCGC DYLKQTGLDLHDIINAPK

ATTGAGAACATTACTCTTTGAATGTGGTATAGGTGAAGAAGATATCGACAAAAA LSALRTLLFECGIGEEDID

AGCAAGCCAAGATCAGAATTTTCCTATTCAAAATGTCATATCACAACACAGAGC KKASQDQNFPIQNVISQH

CCTGATTTTCTGCCAACTAAAAGATATGCTAGACATGGTTGAAAATGACTTGTTl RALIFCQLKDMLDMVEN

AAAAAGTATATGCCCTCCGTAACCTATATGAGGCTAGATGGAAGCATTGACCCA DLFKKYMPSVTYMRLDG

AGAGACAGACAAAAAGTTGTTCGGAAATTTAACGAAGATCCCTCTATTGATTGC SIDPRDRQKWRKFNED

CTACTGTTGACCACCAAGGTCGGAGGGCTGGGTCTGAATTTAACTGGTGCAGA PSIDCLLLTTKVGGLGLN

CACCGTCATTTTTGTAGAGCATGACTGGAATCCAATGAATGATCTACAGGCAAT LTGADTVIFVEHDWNPM

GGACAGAGCACATAGAATTGGTCAGAAAAAGGTTGTTAATGTTTACAGAATTAT NDLQAMDRAHRIGQKKV

TACGAAAGGTACGCTTGAAGAAAAAATCATGGGTTTGCAGAAATTCAAAATGAA VNVYRIITKGTLEEKIMGL

TATAGCTTCTACAGTTGTTAATCAGCAAAACAGTGGATTAGCATCCATGGATAC QKFKMNIASTWNQQNS

ACATCAGCTGCTTGATCTCTTCGACCCCGATAACGTTACCTCACAGGACAATGA! GLASMDTHQLLDLFDPD

GGAAAAGAATAATGGCGATTCCCAAGCAGCCAAGGGCATGGAAGATATTGC NVTSQDNEEKNNGDSQ

AAKGMEDI

YGL10 365 ATGGGCCCCGCTGTCCCTGCGATGCCTCCAGTACCATCAAATTTTCCGCCTGT 366 MGPAVPAMPPVPSNFPP 8c TCCAACTGGTACAATAATGTCGCCTCAGTTGAGCCCTTTTCCGGATCACCGTTT VPTGTIMSPQLSPFPDH

AAGACATCACCCATTAGCTCATATGATGCCTGCTGATAAGAATTTTCTGGCATA RLRHHPLAHMMPADKNF

TAACATGGAGTCTTTCAAAAGTAGAGTGACTAAAGCATGTGATTATTGTCGGAA LAYNMESFKSRVTKACD

GAGGAAGATTAGATGTACGGAAATCGAGCCGATTTCTGGTAAATGTAGAAACT YCRKRKIRCTEIEPISGK

GTATCAAGTATAATAAAGATTGCACGTTTCATTTCCATGAAGAACTGAAAAGAA CRNCIKYNKDCTFHFHE

GGCGAGAAGAAGCTTTGAATAACAAGGGAAATGGGAAGTCGGTTAAGAAGCC ELKRRREEALNNKGNGK

GAGGTTGGATAAGGAGAATAAGTTCAAGGATGAAAATTTTGATATCGCCGTGC SVKKPRLDKENKFKDEN

GATCAAGAAATACTTCTTCTACAGACAGCTCGCCGAAATTACACACCAACTTAT FDIAVRSRNTSSTDSSPK

CACAAGAATATATTGGAGTTTCTGCCGGCAAAAGTGCAAGCGATAAAGAAGAT LHTNLSQEYIGVSAGKSA

ACTTGGCCTGACTTTGTTCCTATTGACAGGACAGTGCTTGAAAAAATTGAACTC SDKEDTWPDFVPIDRTV

AACCATACGAAGGTTGCTGGGAAAGTTTTTGTTTTGGAAGAGATTTGTAAAAAC LEKIELNHTKVAGKVFVL

ATGAAGGGAACAATAGAGAAGCTAGCTGAAAAAAGCAAAATTGACGTCATTGATI EEICKNMKGTIEKLAEKS

AAAGAATACATGAAAAGGCCTAAAAGAAAGCAATATTCAAAGGCCTTGTTAACA KIDVIDKEYMKRPKRKQY

AAACAAAAAATGTTTCATTTTCGACAAAATGTTTTATCACATTTAACTGACGAAG SKALLTKQKMFHFRQNV

YOL12 41 - TTGACAATCGATGAGAAGGAAATTTTGAAAGGATGTAATGAGGAAATAAAAATT 412 LTIDEKEILKGCNEEIKIKL

8c AAGCTGGAAAGGTTGAATGAAAGACTAGGATCATGGGAAAAAAGCAAAGAAAA ERLNERLGSWEKSKEKY

ATACGAAACATCATTGAAGGATAAAGAAAAAATGCTAGCAGATGCTGAAAAGAA ETSLKDKEKMLADAEKK

AACAAACACTCTATCAAAGGAACTGGATAATTTGAGGTCGCGATTTGGAAACTT TNTLSKELDNLRSRFGNL

AGAGGGAAATACTTCTGAAAGGATTACAATAAAAAATATCTTACAGTCAAGGCC EGNTSERITIKNILQSRPD

CGATATTTCGGCAGAAGAATGTAATTTCCTTATGGTTGAACAAATTGATTCAGC ISAEECNFLMVEQIDSAN

AAATTTGACGACTTTACAAAATACTGTCAAAGAAATTGTTTTGGCTGTTGGTATT LTTLQNTVKEIVLAVGIPY

CCTTATCCAAAGTTAAGGCGAAAGATTCCGTTGCTGGCTATCAAATTGAAATAT PKLRRKIPLLAIKLKYENI

GAAAACATCATGCTGTCCAATTTTGCTCAACGTCTGCATAGACAAGTATATAGT MLSNFAQRLHRQVYSQE

CAAGAAATGAATCTAAAGAAATTTACGGATCAAGCATACTATGATTTTATGTCAA MNLKKFTDQAYYDFMST

CGCGCAGGATGGATTCCATAGATCACCATTTGGAGAGGTGTCTGGACCATCTG RRMDSIDHHLERCLDHL

TATGATCATATCCTGGAGAAGATGGTGAAGTAATTTTTATTAACGGAGTCAATA YDHILEKMVK

GATAAATTGTTGTAATTGTAACTAAAAATTAAGGTATAAGATTAAATTATATTCTA

GGAAGTATGTATAAAATATTCATTTGTTGTAAAATTGGAGGGCTTAAGCGGCCA

ACATTTTATGTTTACACTTTTTCTTGTTTTAAGAATAGAAAAAAAATATACGTGAA

AAAAATTTGAAGTCATCGCAAAGTAGCTCTTTTTACATAGTAAAAGTCTGACCA

GCCAAGAGTTAATATCAGTATAGTGATATCCTACTACTTCAAAAATAACGGATG

AAGACCTGCTACTATG

YOL12 413 GACGATAGATCAACATCACCAAAATCGGCAATCGAACTATATCAAAGATTTCAA 414 DDRSTSPKSAIELYQRFQ 8c CAGATGATTAAGGAACTAGAGCTGAGTTTTGACGCAAGTCCTTACGCAAAATAC QMIKELELSFDASPYAKY

TTCCGCCGGTTGGATGGAAGGCTTTGGCAAATAAAGACAGACTCAGAATTAGA FRRLDGRLWQIKTDSEL

AAACGATGAATTGTGGCGATTAGTCTCAATGAGCATATTTACAGTATTCGATCC ENDELWRLVSMSIFTVF

TCAGACCGGCCAAATTCTAACTCAAGGACGCAGGAAGGGAAACTCCTTAAATA DPQTGQILTQGRRKGNS

CATCAACTAAAGGCTCCCCATCAGATTTACAGGGAATAAACAACGGGAACAATAI LNTSTKGSPSDLQGINN

ATGGGAACAATGGTAATATTGGAAATGGGAGTAATATTAAGAACTATGGAAATA GNNNGNNGNIGNGSNIK

AAAACATGCCAAACAACCGAACGAAAAAAAGAGGCACCAGGGTGGCTAAAAAT NYGNKNMPNNRTKKRG

GCTAAAAATGGGAAAAACAATAAAAATAGTAATAAAGAGAGAAACGGCATTACA TRVAKNAKNGKNNKNSN

GATACGAGTGCATTCAGTAATACAACAATAAGCAACCCAGGTACCAATATGCTT KERNGITDTSAFSNTTIS

TTTGATCCATCATTGTCTCAACAGTTACAAAAACGACTGCAAACGCTATCACAA NPGTNMLFDPSLSQQLQ

GATGTCAATTCTCGTTCGTTGACAGGATATTATACACAGCCAACCAGTCCTGGC KRLQTLSQDVNSRSLTG

TCAGGAGGATTTGAATTTGGTTTGAGTCATGCAGATCTGAACCCCAATG YYTQPTSPGSGGFEFGL

SHADLNPN

YOL12 415 lCCGAAAGTTGGAAACTGCTACTTCAGAAACAAGAGAGGGTATTTCTTCGGAGC 416 RKLETATSETREGISSEL 8c TTTCCTCATTTTTAAACGGGAACATCATCGAGCATGACGTTCCCGAAGTTTTTTT SSFLNGNIIEHDVPEVFF TGATGAATTTCAAAAAGCTATCCAGAGTAAACAAAAGGCGCTCAACACGCTGG DEFQKAIQSKQKALNTLG GTGCGGTGGCTTACATAGCAAATGAAACTAATCTATCACCTTCTGTTGAACCGT AVAYIANETNLSPSVEPYI ACATAGTTGCGACGGTCCCTTCTGTATGCAGTAAAGCTGGTAGTAAAGATAATGl VATVPSVCSKAGSKDND ATGTTCAACTTGCGGCAACAAAGGCCCTCAAAGCCATCGCTAGTGCTGTTAAC VQLAATKALKAIASAVNP CCGGTTGCCGTTAAAGCACTCTTACCTCATTTGATTCATTCGCTGGAAACTAGC VAVKALLPHLIHSLETSN AACAAGTGGAAAGAGAAGGTTGCTGTTCTCGAAGTAATCTCTGTATTGGTAGAT KWKEKVAVLEVISVLVDA GCCGCTAAAGAACAGATTGCCTTAAGAATGCCGGAACTAATTCCCGTTCTTTCG AKEQIALRMPELIPVLSE GAATCCATGTGGGACACTAAGAAAGGAGTTAAGGAGGCGGCAACGACAACCA SMWDTKKGVKEAATTTI TCACAAAGGCAACTGAAAC TKATE

YOL12 417 GAAGCGGAACCTTATGATAGTGATGAGGCAATCTCTAAGATTTCCAAGAAAAAG41 8 EAEPYDSDEAISKISKKK 8c ACTAAGAAAATAAAGACCGAACCAGTGCAATCGTCGTCATTACCATCGCCTCCA TKKIKTEPVQSSSLPSPP

GCAAAGAAAAGCGCGACATCAAAGCCTAAAAAAATCAAGAAAGAAGATGGTGA AKKSATSKPKKIKKEDGD

TGTAAAGGTAAAAACAACTAAAAAGGAAGAACAGGAGAACGAAAAAAAGAAAC VKVKTTKKEEQENEKKK

GAGAAGAAGAAGAAGAGGAGGACAAGAAAGCGAAGGAGGAGGAGGAAGAATA REEEEEEDKKAKEEEEE

TAAATGGTGGGAAAAAGAAAACGAAGATGACACCATAAAATGGGTCACACTGA YKWWEKENEDDTIKWVη

AGCATAACGGTGTTATATTCCCTCCACCATACCAGCCCTTACCATCTCACATCA LKHNGVIFPPPYQPLPSH

AATTATATTACGATGGGAAGCCAGTAGATTTACCTCCGCAAGCTGAAGAAGTAGl IKLYYDGKPVDLPPQAEE

CCGGGTTCTTTGCTGCCCTATTAGAGAGTGATCATGCCAAAAATCCTGTTTTCC VAGFFAALLESDHAKNP

AAAAGAACTTCTTCAATGATTTCTTGCAAGTACTGAAAGAAAGTGGTGGTCCCC VFQKNFFNDFLQVLKES

TCAATGGAATTGAGATAAAGGAATTTTCTCGTTGCGATTTCACCAAAATGTTTGA¹ GGPLNGIEIKEFSRCDFT

TTACTTCCAGTTACAAAAAGAACAGAAAAAGCAACTGACTTCCCAAGAAAAGAA KMFDYFQLQKEQKKQLT

ACAGATTCGTTTGGAAAGAGAAAAATTCGAGGAAGATTATAAATTCTGTGA SQEKKQIRLEREKFEEDY

KFC

YOL12 419 GTCAATTTCAAAGTACTTCACTCCCGTTGCTGACGGGTCACTCACTTTCAATGG 420 SISKYFTPVADGSLTFNG 8c CGCGAACATTCAATTTGGCGCCGATGCTCAAGGCGAGTCAAAAAAGAGTTATG ANIQFGADAQGESKKSY

ACGCTGAGGACAGCATGCCGAATCCGGCAAATCAACTAAATGACATAACCTTC DAEDSMPNPANQLNDIT

CAAGCAGAGGCTGGTGAAATGGTTTTGGTTTTGGGTTATCCCACATCCACTCTA FQ AEAG EMVL VLG YPTS

TTTAAGACTTTGTTTCATGGTAAAACTAGTTTGTCATACTCTCCTCCAGGCTCGA TLFKTLFHGKTSLSYSPP

TTAAATTTAAAAATAATGAGTTTAAGAGCTTTTCCGAAAAATGTCCCCACCAAAT GSIKFKNNEFKSFSEKCP

CATTTACAAT HQIIYN

YOL12 421 GTGCTTTAGAGATGAAGATTTAAGAGCAGACAGGCAGCCTGAGTTTACACAGG 422 CFRDEDLRADRQPEFTQ 8c TTGATATGGAAATGGCCTTTGCTAATTCTGAAGATGTCATGAAAATCATAGAAA VDMEMAFANSEDVMKIIE

AGACAGTTTCTGGGGTATGGAGTAAATTTTCCAAAAAACGAGGATTATTGACTT KTVSGVWSKFSKKRGLL

TAGACAGTAAGGGTACATTAGTGCCTGCGAAAAAGGAAAACGGCACAGTATCT TLDSKGTLVPAKKENGT

ATCTTTCGTATGACCTACGAACAAGCCATGACCTCATATGGTATTGACAAGCCA VS I FRMTYEQAMTS YGI D

GATTTGAGAGCGCCAGATTTGAAGATTATCAATTTAGGCGAGTTCAATGCCTTT KPDLRAPDLKIINLGEFN

AGTCATTTGAACAAAAAATTTCCCGTTTTTGAAGTAATTATTCTAAGAAGTGCCT AFSHLNKKFPVFEVIILRS

TTTCAAATATGGAAGAGTACAAAGAACGATGGTCGTTTCTGACAAATAACAGTA AFSNMEEYKERWSFLTN

ATTACAATTATAGAGTTCCAATAGTGCTACCAATTGAAAATGACGAACAAGCTA NSNYNYRVPIVLPIENDE

ATTCAAATTGGTTTGAGAATTTTCATGCAATTGCCACGTTTGAAAACCCACATCT QANSNWFENFHAIATFE

AATAACCAAATTTCTGAAACTGAAAAAAGGTGACATTGTATGCGGTTGTACGAG NPHLITKFLKLKKGDIVC

AGAGCCAAACCATTCCATTTTCGAGAATCCTACTCCCCTGGGAAGATTGAGAC GCTREPNHSIFENPTPLG

AGTTGGTGCTACAAAGTGAGCATGGGAAAAATATCTATCATGCTGTCAATAAGG RLRQLVLQSEHGKNIYH

ATGTTGCCTCATGGATTGTGGATTTCCCGTTATTTTCTCCCGTTATAATTGAAGA AVNKDVASWIVDFPLFSP

TAAGTCTGGTAAAAAAGAAAAGCTTGCATATCCGGAGTACGAAAAGGATAGACT VIIEDKSGKKEKLAYPEY

ATGTTCCACGCATCATCCTTTTACTATGGTGAAGCTTAAAG EKDRLCSTHHPFTMVKL

K

YPL14 423 CAAAAAAGCATGACTTCTTCGCCTCTGAAAAATGTTCTACCTGATCTTAAAGAATI424 QKSMTSSPLKNVLPDLK 1c CGTCTCCCTTAAACGATAGTAGGGAAGACACAGAATCAATAACATACTCATACG ESSPLNDSREDTESITYS

ATTCCGAGTTGTCATCCAGCTCTCCACCCAGAGATACTGTAACGAAAAAGTCAA YDSELSSSSPPRDTVTK

GAAAAGTCAGAAACATCGTGAATAATACAGATAGTCCAACTCTAAAAACAAAAA KSRKVRNIVNNTDSPTLK

CTGGATTTTTAAATCTAAGAGAGTTTACATTTGAAGATACCAAGTCTTTAGATGA TKTGFLNLREFTFEDTKS

GAAAAAGAGTACTATAGATGGCCTTGAAAAGAATTATGATAACAAAGAAAATCA LDEKKSTIDGLEKNYDNK

GGAATCAGAATATGAGAGCACAAAAAAGCTGGACAACTCACTCGATGCATCAT ENQESEYESTKKLDNSL

CAGAAGCCAATAATTATGATATAACTACAAGAAAAAAACATTCATCTTGCAATCA DASSEANNYDITTRKKHS

CAAGATCAAACAAGCAGTTGTAAGACCGGCTAGTGGAAGGATCAGCATTTCAA SCNHKIKQAWRPASGRI

GAGTTCAAAGCATTGCCATAACACCAACCAAAGAG SISRVQSIAITPTKE

YPL14 425 CATGGATACCACGATGGATCCACCCTCTTCCATGAATGATGCGTTAAGAGAAG 426 MDTTMDPPSSMNDALR 1c TTGTCGAGGATGAAACTGAGTTATTTCCTCCTAATTTAACTAGACGCTATTTCCT EWEDETELFPPNLTRRY

TTATTTTAAGCCTCTATCGCAAAATTGTGCTCGTCGTTACAGGAAGAAAGCAAT FLYFKPLSQNCARRYRK

TAGTTCTAAACCATTATCTGTTAGGCAGATTAAAGGTGACTTCCTAGGCCAATT KAISSKPLSVRQIKGDFL

GATTACCGTCAGAGGTATTATCACCAGAGTTTCTGATGTCAAACCAGCTGTGGA GQLITVRG I ITRVS DVKPA

AGTTATCGCATATACCTGCGATCAATGTGGGTACGAAGTTTTCCAAGAGGTCAA VEVIAYTCDQCGYEVFQ

CTCTCGTACTTTTACTCCGTTGTCAGAATGTACTTCCGAAGAATGTTCCCAAAA EVNSRTFTPLSECTSEE

YPL1 1431 TCAATACTCTTCGTTTTTGGCGCCCTTGGCAGTGGACTCCGTATTGAAGATATC 432 QYSSFLAPLAVDSVLKIS 1c TGATGAAAACTCTAAGAATGTTGACCTGAACGATATCAGACTGGTCAAAAAAGT DENSKNVDLNDIRLVKKV

TGGTGGTACCATTGATGACACAGAAATGATAGATGGTGTGGTCTTGACACAAA GGTIDDTEMIDGWLTQT

CGGCAATCAAATCTGCTGGTGGTCCGACAAGAAAGGAAAAAGCAAAGATTGGG AIKSAGGPTRKEKAKIGLI

TTAATTCAATTCCAAATATCTCCTCCAAAGCCCGACACAGAAAATAATATCATCG QFQISPPKPDTENNIIVN

TTAATGACTATAGACAAATGGATAAGATCCTTAAAGAAGAAAGAGCGTATTTGC DYRQMDKILKEERAYLLN

TAAATATCTGTAAAAAAATTAAAAAGGCCAAGTGTAACGTGCTGTTGATTCAGA ICKKIKKAKCNVLLIQKSIL

AATCCATCTTGAGAGATGCGGTAAATGATTTGGCTCTTCATTTCTTGTCAAAATT RDAVNDLALHFLSKLNIM

GAACATAATGGTGGTAAAGGATATCGAGAGAGAAGAAATCGAGTTTCTGTCGA VVKDIEREEIEFLSKGLG

AGGGCTTGGGTTGTAAGCCAATTGCTGATATAGAATTGTTCACCGAAGATAGAT CKPIADIELFTEDRLGSA

TGGGTTCCGCTGATTTAGTTGAGGAGATCGATAGC DLVEEIDS

YPL14 433 GATATATGTCTCTCCCCAGCAGGATTTTTCTGACTTGATGAAGTCTTGGAAAAA 434 IYVSPQQDFSDLMKSWK 1c TGAACGGTGTTCGCCAGAACTCTTACCATATCCTCATCAGTTGATGAAAAGATT NERCSPELLPYPHQLMK

ATTGAATCGAATATCCATGCAATCTCAATTAATTGAAAACATTTCAATGGGATTC RLLNRISMQSQLIENISM

CTCGATATGCAAAATGCTTCTAATGCGAACCCGCCTATGCCTAACGAATCTAAA GFLDMQNASNANPPMP

TTGCCTTTACTCTGCATGGAAACTGAGTTAGAAAGATTGAAATTTGTTATTAGAA NESKLPLLCMETELERLK

GCTACATACGATGTAGGCTAAGCAAAATTGACAAATTCTCACTTTATTTGCGCC FVIRSYIRCRLSKIDKFSL

AATTAAATGAAGATGAAAATTCGCTGATCTCTCTTACAGATTTACTATCCAAAGA YLRQLNEDENSLISLTDL

TGAAATCAAATACCATGACACGCATTCCCTGATCTGGTTGAAACTAGTCAATGA LSKDEIKYHDTHSLIWLK

TTCTATCCTGAAATACATGCCCGAAGAACTTCAGGCTATTAATGATACAGAAGG LVNDSILKYMPEELQAIN

TAGCGTAAACATGATAGATGAGCCCGACTGGAACAAATTTGTCTTTATACACGT DTEGSVNMIDEPDWNKF

TAACGGCC VFIHVNG

YPL14 435 GGCAGCAAAAAGCATCTTAAACAGTTATCAGATGAGGAAGAACGAATGAGAGA 436 GSKKHLKQLSDEEERMR 1c AAAAATGTCCATACGCAAAGCTAGTGCTCTCGAGTGGGAAAGATTTTTGCTTAC EKMSIRKASALEWERFLL

CGACTTCAGTGCGATAATTGACCTTTTCCAAGGACAGTACGCATCTAGGCTACA! TDFSAIIDLFQGQYASRL

ATGTCAAGTTTGTGAACATACCTCCACAACTTACCAAACATTCTCTGTTCTTTCT QCQVCEHTSTTYQTFSV

GTTCCTGTCCCACGCGTTAAAACTTGTAACATATTAGACTGTTTCCGGGAATTC LSVPVPRVKTCNILDCFR

ACCAAATGCGAAAGGTTAGGTGTCGATGAACAATGGTCATGTCCTAAATGCTTA EFTKCERLGVDEQWSCP

AAAAAGCAGCCTTCCACTAAACAACTGAAGATTACTAGATTGCCTAAGAAACTA KCLKKQPSTKQLKITRLP

ATTATTAATTTGAAACGATTTGACAATCAAATGAATAAGAATAATGTGTTTGTCC KKLIINLKRFDNQMNKNN

AATATCCTTACTCCTTAGATCTTACACCATATTGGGCGAGAGATTTTAATCATGA VFVQYPYSLDLTPYWAR

AGCTATTGTTAATGAGGACATTCCTACCAGGGGCCAAGTACCACCATTTAGATA DFNHEAIVNEDIPTRGQV

CAGACTGTATGGGGTTGCATGTCATTCGGGGAGTTTGTATGGGGGACACTATA PPFRYRLYGVACHSGSL

CTTCATACGTTTATAAGGGACCAAAAAAAGGTTGGTATTTTTTCGATGACTCGC YGGHYTSYVYKGPKKG

CGCTTGGGTCACTCAAGGAAAGGAATTGCATGAAAAAGGTTGGGTAGGGGAT LVAWVTQGKELHEKGW

GCCAAAACTGGCGATTTGCAAGAACAGTTCAATATAGCCACTGTCAAATTATAT VGDAKTGDLQEQFNIAT

GAAAGCGCAGAAGACGGGCGTCTTTCCATTGGTAAAGACAAACCCTTTCGAGA VKLYESAEDGRLSIGKDK

GGAAAACACCGGCAGTGATAGTTTGGTTCGAGCGGAAGAGGACTCTACCGCT PFREENTGSDSLVRAEE

GTCACTAAT DSTAVTN

YDR21 453 GCCATATTCGGAGGCG1 [ΥCTTAGGGTTCAATAACCCAACGCCTGGATTAGA454 PYSEAFFLGFNNPTPGL 9c AGCTGAGCACTCAAGCACATCGCCTGCCCCCGAGAACTCCGAAACACATAATA EAEHSSTSPAPENSETH

GGAAAAGAAATAGAATATTGTTTGTCTGCCAGGCTTGTTGGAAGTCAAAAACAA NRKRNRILFVCQACWKS

AGTGTGATAGAGAAAAACCTGAATGTGGTCGATGCGTCAAGCATGGGTTAAAA KTKCDREKPECGRCVKH

TGTGTTTATGACGTATCAAAACAGCCAGCACCACGAATTCCGAGTAAAGACGC GLKCVYDVSKQPAPRIP

CATTATATCAAGGTTGGAAAAAGATATGTTTTATTGGAAAGATAAAGCTATGAAG SKDAIISRLEKDMFYWKD

CTACTAACAGAGAGAGAGGTGAATGAATCAGGCAAGAGATCAGCAAGTCCGAT KAMKLLTEREVNESGKR

CAATACAAACAATGCTAGCGGGGACAGTCCTGATACCAAGAAGCAGCATAAAA SASPINTNNASGDSPDT

TGGAACCTATATATGAACAAAGTGGTAACGGGGATATAAACAATGGTACCAGAA KKQHKMEPIYEQSGNGD

ATGATATTGAAATCAACTTGTATAGAAGTCATCCAACCATGATCATGAGTAAAGT INNGTRNDIEINLYRSHPT

CATGAAAAGAGAAGTTAAGCCGTTATCTGAAAATTATATTATAATTCAGGACTGT MIMSKVMKREVKPLSEN

TTTCTAAAAATCCTGGTCACTTCAGTGTTCCTTGACACTTCAAAGAACACGATG YIIIQDCFLKILVTSVFLDT

ATACCGGCATTGACGGCAAACGCGAATATTACAAGAGCCCAGCCTAGCGT SKNTMIPALTANANITRA

QPS

YDR21 455 AATCACGAGAGTGAGCCTGGATGAAGCATTACCCAAAACGTTTTATGACATGTA456 ITRVSLDEALPKTFYDMY 9c TTCGCCAGATATTCTATTAGCAGACCCATCCAACATTCTCTGTAACGGGCGTCC SPDILLADPSNILCNGRP

CAAGTTTACCAAGAGAGAGTTATTGGATTGGGATTTAAACGATATAAGATCGTT KFTKRELLDWDLNDI RSL

ATTGATAGTCGAGAAGTTAAGGCCCGAATGGGGTAATCAACTACCGGAAGTAA LIVEKLRPEWGNQLPEVI

TAACGGTGGGTGATAATATGCCCCAGTTTAGGTTACAATTATTACCACTATATT TVGDNMPQFRLQLLPLY

CTAGCGATGAGACCATAATCGCAACGTTAGTCCATTCGGATCTGTACATGGAG SSDETIIATLVHSDLYME

GCTAACTTAGATTATGAATTCAAACTAACCAGCGCCAAATATACAGTAGCGACC ANLDYEFKLTSAKYTVAT

GCTAGAAAAAGACATGAGCATATAACTGGTAGAAATGAAGCCGTCATGAATTTG ARKRHEHITGRNEAVMN

TCGAAACCGGAATGGAGAAATATCATCGAAAATTACCTCTTAAATATAGCAGTA LSKPEWRNllENYLLNIAV

GAGGCACAATGCAGGTTTGATTTCAAACAAAGATGCTCCGAATATAAGAAATGG EAQCRFDFKQRCSEYKK

AAGTTACAACAGTCCAACTTAAAAAGACCGGACATGCCCCCACCAAGCATAATA WKLQQSNLKRPDMPPP

CCGCGGAAAAACAGCACAGAAACAAAATCGCTTCTGAAAAAGGCTTTATTGAA SIIPRKNSTETKSLLKKAL

GAACATTCAGTTGAAAAACCCCAATAATAACCTTGATGAATTGATGATGAGATC LKNIQLKNPNNNLDELM

AAGCGCCGCAACAAATCAACAGGGA MRSSAATNQQG

YDR32 511 AAGTCAGAGGCGCCAAGACCACATGCTTGTCCTATCTGTCATAGAGCTTTTCA 512 KSEAPRPHACPICHRAF 8c CAGACTGGAACATCAGACGAGACACATGAGAATTCATACAGGTGAGAAGCCTC HRLEHQTRHMRIHTGEK

ACGCGTGTGACTTCCCCGGATGTGTGAAAAGGTTCAGTAGAAGCGATGAACTG PHACDFPGCVKRFSRSD

ACGAGACACAGAAGAATTCATACAAACTCCCACCCTCGAGGTAAAAGAGGCAG ELTRHRRIHTNSHPRGK

AAAGAAGAAGGTTGTGGGCTCTCCAATAAATAGTGCTAGTTCTAGTGCTACCA RGRKKKVVGSPINSASS

GTATACCAGATTTAAATACGGCAAATTTTTCACCGCCATTACCACAGCAACACC SATSIPDLNTANFSPPLP

TATCGCCTTTAATTCCTATTGCTATTGCTCCGAAAGAAAATTCAAGTCGATCTTC QQHLSPLIPIAIAPKENSS

TACAAGAAAAGGTAGAAAAACCAAATTCGAAATCGGCGAAAGTGGTGGGAATG RSSTRKGRKTKFEIGES

ACCCATATATGGTTTCTTCTCCCAAAACGATGGCTAAGATTCCCGTCTCGGTGA GGNDPYMVSSPKTMAKI

AGCCTCCACCTTCTTTAGCACTGAATAATATGAACTACCAAACTTCATCCGCTT PVSVKPPPSLALNNMNY

CCACTGCTTTGTCTTCGTTGAGCAATAGCCATAGTGGCAGTAGACTGAAACTG QTSSASTALSSLSNSHS

AACGCGTTATCGTCCCTA GSRLKLNALSSL

YDR32 513 CGTCAGACAACCCTCCCTAGGACCCACTTTAGGTGTCAAAGGTGGTGCTGCG 514 VRQPSLGPTLGVKGGAA 8c GGTGGTGGTTATTCCCAAGTCATCCCAATGGACGAATTCAACTTACATTTGACT GGGYSQVIPMDEFNLHL

GGTGACATTCACGCCATTGGTGCGGCTAACAACCTACTTGCTGCCGCTATTGA TGDIHAIGAANNLLAAAID

CACTAGAATGTTCCATGAGACCACTCAAAAGAACGACGCTACCTTCTACAACAGI TRMFHETTQKNDATFYN

ACTAGTGCCTAGAAAGAACGGAAAGAGAAAGTTTACTCCCTCCATGCAAAGAA RLVPRKNGKRKFTPSMQ

GATTGAACAGACTGGGTATTCAAAAGACCAACCCCGATGATCTAACACCCGAA RRLNRLGIQKTNPDDLTP

GAGATCAACAAATTCGCCAGATTGAACATTGACCCGGACACTATTACTATCAAG EEINKFARLNIDPDTITIKR

AGGGTGGTCGATATCAACGACAGAATGTTAAGACAAATCACCATTGGTCAAGC VVDINDRMLRQITIGQAP

CCCTACCGAGAAGAACCACACAAGAGTTACTGGATTCGATATCACCGTTGCTT TEKNHTRVTGFDITVASE

CTGAATTGATGGCTATTCTTGCTCTTTCAAAGGACTTGAGGGACATGAAGGAAC LMAILALSKDLRDMKERI

GTATTGGAAGAGTCGTTGTTGCTGCTGACGTAAACAGGTCTCCAGTCACTGTT GRVVVAADVNRSPVTVE

GAAGATGTGGGTTGTACCGGTGCCTTAACCGCTTTATTAAGAGACGCTATCAA DVGCTGALTALLRDAIKP

GCCCAACTTGATGCAAACTTTAGAAGGTACTCCTGTCTTGGTCCATGCCGGCC NLMQTLEGTPVLVHAGP

CATTTGCCAACATCTCTATCGGTGCCTCTTCTGTTATTGCTGATCGCGTGGCTT FANISIGASSVIADRVALK

TGAAATTGGTTGGTACCGAGCCAGAGGCAAAAAC LVGTEPEAK

ATGTTTTGGATAAAGATGCGTTTATTCCAAAATTGAAACCCGCACCTATCAACA GDVLDKDAFIPKLKPAPI

GTTTAAGTCGTGATTTCGTGATGAAAACAAGAAGGCGGAAGGGTATTTCTACA NSLSRDFVMKTRRRKGI

GGTGGATTTATGTCAAATGATGGTCCTACGCTTGAAAAGTATATAAGCGCTGAA STGGFMSNDGPTLEKYI

TTATACGCTCAATTAAGAGAAAATGGCTTAGTACCGTGAAAATCTTGGCGCATA SAELYAQLRENGLVP

AGAATTAAGCAATATGTCCACAATATTTTTATTCAGCAGAATTTACATATGACTT

CG

YDR32 527 TTGTTTCACTGTCAAAACGCCCCTATGGAGAAGAGATGAACACGGTACTGTTCT 528 CFTVKTPLWRRDEHGTV 8c CTGTAATGCATGTGGCCTCTTCCTGAAGTTGCACGGGGAACCAAGGCCTATCA LCNACGLFLKLHGEPRPI

GCTTGAAGACGGACACCATTAAGTCAAGAAATAGGAAAAAGCTGAATAACAAC SLKTDTIKSRNRKKLNNN

AATGTGAACACTAATGCCAATACCCATTCTAACGACCCAAATAAAATATTCAAG NVNTNANTHSNDPNKIF

AGAAAGAAGAGACTGCTTACAACTGGTGGTGGTTCATTACCTACGAATAATCC KRKKRLLTTGGGSLPTN

GAAGGTTTCTATTCTGGAAAAGTTTATGGTGAGCGGGTCCATTAAGCCACTGTT NPKVSILEKFMVSGSIKP

AAAACCAAAGGAAACCGTTCCCAACACAAAGGAGTGCTCCACGCAGCGGGGA LLKPKETVPNTKECSTQ

AA RG

YDR32 529 GCGCGCGGAAGTGTTCCTCTAAAGTCGGTTGGCAGTGGATTAACAAAGAAAGC 530 ARGSVPLKSVGSGLTKK 8c AACAACATCAATAACAAGTAATTCAGCTACCACAACATTTGAACGACAGTACCT ATTSITSNSATTTFERQY

TATAAAGTATCTTTATCGGCACCAGGCCTATGGGAACGTTATAAAAATTGCACA LIKYLYRHQAYGNVIKIAQ

GAAGTTTCTTTATACCACTATTGGTTCACAAAGATTGTTAAAACAGGATGCCTCAi KFLYTTIGSQRLLKQDAS

TTACCTGAATTGAAAAAGTTCCTTCTCTCTTTATTGATTTTACAAAGAGGTATTC LPELKKFLLSLLILQRGIQ

AATTAGATCAGGCAATCTCTGATATCATACAACGGTTTCTATTAACACAAAAGAC LDQAISDIIQRFLLTQKTM

AATGGTGATAGACCTCATTAACTCGATTTTCTCTAGGATGGTTATAATGAATATG VIDLINSIFSRMVIMNMHE

CATGAAGAAGCTGTATATAAATGGGTCAAATGGATGAAACTAGTGAATGGACAT EAVYKWVKWMKLVNGH

TGTGAATTTACGAATTATATGGAGAACAAGATAGTTTTGAGAAACTTTTTATCAT CEFTNYMENKIVLRNFLS

TCATGAGGCAATCAAATGTTCGCCCCGATTATTTATCTTATTTGAAAGCAATTCA FMRQSNVRPDYLSYLKAI

GCTAACGCAAGGGCCCGCAATAGCGTCTCAATTTGCAACTACGTTGTTGTTCTT QLTQGPAIASQFATTLLF

ATTAACTTATATTAGGAAATTTTCTTCTGCAGAAGCAGTTTGGAATTACAAGTGC LLTYIRKFSSAEAVWNYK

GAACATAACCTGCCGATAGTGAGTTCTGACCTGACGTGTATTCTAAAGACGTAT CEHNLPIVSSDLTCILKTY

TGTCACATGCAGAAGTTTAATTTGGTTTCGTCAACATATTGGAAATACCCTGAT CHMQKFNLVSSTYWKYP

YDR32 535 GCGAGCTCTAGTGCCACCTCAAATGGAATATATACGCAAGCGCAATATTCTCAA536 ASSSATSNGIYTQAQYS 8c CTTTTCGCCAAAATATCAAAACTATATAACGCTACACTATCATCTGGGTCAATTG QLFAKISKLYNATLSSGSI

ACGATAGATCAACATCACCAAAATCGGCAATCGAACTATATCAAAGATTTCAAC DDRSTSPKSAIELYQRFQ

AGATGATTAAGGAACTAGAGCTGAGTTTTGACGCAAGTCCTTACGCAAAATACT Q IKELELSFDASPYAKY

TCCGCCGGTTGGATGGAAGGCTTTGGCAAATAAAGACAGACTCAGAATTAGAA FRRLDGRLWQIKTDSEL

AACGATGAATTGTGGCGATTAGTCTCAATGAGCATATTTACAGTATTCGATCCT ENDELWRLVSMSIFTVF

CAGACCGGCCAAATTCTAACTCAAGGACGCAGGAAGGGAAACTCCTTAAATAC DPQTGQILTQGRRKGNS

ATCAACTAAAGGCTCCCCATCAGATTTACAGGGAATAAACAACGGGAACAATAA LNTSTKGSPSDLQGINN

TGGGAACAATGGTAATATTGGAAATGGGAGTAATATTAAGAACTATGGAAATAA GNNNGNNGNIGNGSNIK

AAACATGCCAAACAACCGAACGAAAAAAAGAGGCACCAGGGTGGCTAAAAATG NYGNKNMPNNRTKKRG

CTAAAAATGGGAAAAACAATAAAAATAGTAATAAAGAGAGAAACGGCATTACAG TRVAKNAKNGKNNKNSN

ATACGAGTGCATTCAGTAATACAACAATAAGCAACCCAGGTACCAATATGCTTT KERNGITDTSAFSNTTIS

TTGATCCATCATTGTCTCAACAGTTACAAAAACGACTGCAAACGCTATCACAAG NPGTNMLFDPSLSQQLQ

ATGTCAATTCTCGTTCGTTGACAGGATATTATACACAGCCAACCAGTCCTGGCT KRLQTLSQDVNSRSLTG

CAGGAGGATTTGAATTTGGTTTGAGTCATGCAGATCTGAACCCCAATGCTTCCA YYTQPTSPGSGGFEFGL

GTAATA SHADLNPNASSN

YDR32 537 ACCAGGAACAGAAGCAGGGATCGTTCCGGTTTCAGCAAACACTCCAAAAAGCT 538 PGTEAGIVPVSANTPKSL 8c TGAATAGCAATATTAATATCAACGTAAATAATAACAATATTGGCCAACAGCAAGT NSNININVNNNNIGQQQV

TAAGAAGCCAAGAAAGCAAAGAGTGAAAAAAAAGACCAAAAAGGAATTGGAAC KKPRKQRVKKKTKKELE

TAGAACGTAAAGAAAGGGAGGATTTTCAGAAACGACAACAAAAACTTTTAGAGG LERKEREDFQKRQQKLL

ATCAACAAAGGCAACAGAAATTGCTATTAGAGACAAAATTACGTCAACAATATG EDQQRQQKLLLETKLRQ

AAATCGAACTAAAAAAATTGCCTAAAGTCTACAAGAGATCAATTGTTAGGAACT QYEIELKKLPKVYKRSIV

ACAAACCCCTAATCAACCGCCTCAAGCATTACAATGGTTACGATATCAATTACA RNYKPLINRLKHYNGYDI

TCTCTAAAATAGGAGAGAAAATAGATTCCAACAAGCCAATTTTTCTCTTCGCGC NYISKIGEKIDSNKPIFLFA

CAGAGTTAGGTGCAATTAATTTACATGCTTTATCAATGTCCCTCCAATCGAAGA PELGAINLHALSMSLQSK

ATCTTGGAGAAATAAACACCGCCTTGAACACCTTGTTGGTCACAAGCGCTGAC NLGEINTALNTLLVTSAD

TCGAACTTAAAAATATCTCTGGTCAAATACCCTGAATTATTAGACTCCTTGGCAA SNLKISLVKYPELLDSLAI

TACTCGGCATGAATTTACTGTCAAATTTGTCACAAAATGTTGTTCCATACCATCG LGMNLLSNLSQNVVPYH

AAACACTTCTGACTATTATTATGAGGATGCTGGATCAAATCAATACTATGTTACC RNTSDYYYEDAGSNQYY

CAACACGATAAAATGGTTGATAAAATTTTTGAAAAGGTAAACAACAACGCTACA VTQHDKMVDKIFEKVNN

CTTACACCGAATGATTCTAACGATGAAAAAGTCACTATCCTGGTAGATTCTTTAA NATLTPNDSNDEKVTILV

CAGGTAATCAATTGCCCACCCCTACTCCTACTGAAATGGAGCCTGATCTCGAC DSLTGNQLPTPTPTEME

ACTGAATGTTTTATAAGTATGCAGTCGACATCTCCCGCAGTTAAACAGTGGGAC PDLDTECFISMQSTSPAV

TTATTGCCTGAACCAATAAGATTCCTCCCTAACCAATTTCCTCTGAAAATTCACA KQWDLLPEPIRFLPNQFP

Claims

CLAIMSWhat is claimed is:

1. A complex between two polypeptides of Saccharomyces cerevisiae as defined in columns 1 and 2 of Table I, respectively.

2. A complex between two polynucleotides of Saccharomyces cerevisiae, said polynucleotides encoding two polypeptides as defined in columns 1 and 2 of Table I, respectively.

3. A recombinant host cell expressing a polynucleotide encoding a Saccharomyces cerevisiae polypeptide as defined in columns 1 and 2 of Table I.

4. A method for selecting a modulating compound that inhibits or activates the protein-protein interactions in Table I between two polypeptides of Saccharomyces cerevisiae comprising:

(a) cultivating a recombinant host cell on a selective medium containing a modulating compound and a reporter gene the expression of which is toxic for said recombinant host cell wherein said recombinant host cell is transformed with two vectors:

(i) wherein said first vector comprises a polynucleotide encoding a first hybrid polypeptide and a DNA binding domain;

(iii) wherein said second vector comprises a polynucleotide encoding a second hybrid polypeptide and an activating domain that activates said toxic reporter gene when the first and second hybrid polypeptides interact;

(b) selecting said modulating compound which inhibits the growth of said recombinant host cell.

5. A modulating compound obtained from the method of Claim 4.

6. A SID® polypeptide comprising one of even SEQ ID Nos. 2 to 548 (column 3 of Table II).

7. A SID® polynucleotide comprising one of uneven SEQ ID Nos. 1 to 547 (column 2 of Table II).

8. A vector comprising the SID® polynucleotide of Claim 7.

9. A fragment of said SID® polypeptide according to Claim 6.

10. A variant of said SID® polypeptide according to Claim 6.

11. A fragment of said SID® polynucleotide according to Claim 7.

12. A variant of said SID® polynucleotide according to Claim 7.

13. A vector comprising the SID® polynucleotide according to Claim 11 or 12.

14. A recombinant host cell containing the vector according to Claim 8 or 13.

15. A pharmaceutical composition comprising a modulating compound of Claim 5 and a pharmaceutically acceptable carrier.

16. A pharmaceutical composition comprising a SID® polypeptide of even SEQ ID Nos. 2 to 548 and a pharmaceutically acceptable carrier.

17. A pharmaceutical composition comprising the recombinant host cell of Claim 14 and a pharmaceutically acceptable carrier.

18. A protein chip comprising a polypeptide of Saccharomyces cerevisiae as defined in columns 1 and 2 of Table I .