WO1996027015A2 - A novel gene for a protein kinase associated with drug resistance - Google Patents

Abstract

This invention describes several DNA and amino acid sequences of novel protein kinase, genes and proteins. The principal protein is a 170 kDa kinase. Other embodiments, genes, fragments, proteins, peptides and various segments variants and substitutions are also described. The protein kinase described is associated with multidrug resistance.

Description

A NOVEL GENE FOR A PROTEIN KINASE ASSOCIATED WITH DRUG

RESISTANCE

Field of the Invention

This invention relates to the field of multidrug resistant cells, protein kinases and cancer.

Information Disclosure The fact of the proteins existence was previously known. Sampson, KE., McCroskey, M.C. and Abraham, I. (1993) Identification of a

170 kDa membrane kinase with increased activity in KB-V1 multidrug resistant cells. J. Cell. Biochemistry 52:384-395.

McCroskey, M.C., Sampson, KE., Abraham, I. Isolation and partial characterization of a 170 kDa rat brain membrane associated protein kinase. ASBMB/DBC-ACS Joint Federation Meeting, May 21-25, 1995, abstract.

The DNA that codes for the protein described herein shows some homology to the DNA that codes for a protein found in yeast, the Yak 1 protein.

Garrett, S., Menold, M.M., and Broach, J.R. 1991. The Saccharomyces cer υisiae Yakl gene encodes a protein kinase that is induced by arrest early in the cell cycle. Mol. and Cell. Biol. ll(8):4045-4052. Yakl acts to inhibit cell growth in yeast in a pathway that is parallel and antagonistic to that of PKA(cAMP-dependent protein kinase) (Garrett et al., 1991).

Hartley, A.D., Ward, M.P., and Garrett, S.The Yakl protein kinase of Saccharomyces cereυisiae moderates thermotolerance and inhibits growth by an Sch9 protein kinase-independent mechanism. Genetics 136:465-474, 1994. Over-expression of Yakl induces a thermotolerant phenotype.

Other related and informational references include the following: Ferrell, J.E., Jr., and Martin G.S. Thrombin stimulates the activities on multiple previously unidentified protein kinases in platelets. J. Biol. Chem., _?64:20723-20729, 1989.

Ferrell, J.E., Jr., and Martin, G.S. Identification of a 42-kilodalton phosphotyrosyl protein as a serine (threonine) protein kinase by renaturation. Mol. Cell. Biol., 10:3020-3026, 1990.

Hamada, H., Hagiwara, K-I., Nakεyima, T., and Tsuruo, T. Phosphorylation of the M,. 170,000 to 180,000 glycoprotein specific to multidrug-resistant tumor cells: effects of verapamil, trifluoperazine, and phorbol esters. Cancer Res., 47:2860-2865, 1987.

Massaque, J. Receptors for the TGF-β family. Cell, 69:1067-1070, 1992. Meyers, M.B., Merluzzi, V.J., Spengler, B.A., and Biedler, J.L. Epidermal growth factor receptor is increased in multidrug-resistant Chinese hamster and mouse tumor cells. Proc. Natl. Acad. Sci. USA, 83:5521-5525, 1986.

Meyers, M.B., Shen, W.P.V., Spengler, B.A., Ciccarone, V., O'Brien, J.P., Donner, D.B., Furth, M.E., and Biedler, J.L. Increased epidermal growth factor receptor in multidrug-resistant human neuroblastoma cells. J. Cell. Biochem., 38:87-97, 1988.

Staats, J., Marquardt, D., and Center, M.S. Characterization of a membrane- associated protein kinase of multidrug-resistant HL60 cells which phosphorylates P- glycoprotein. J. Biol. Chem., 265:4084-4090, 1990.

Background Multidrug resistance of tumor cells appears to be a limiting factor in the efficacy of some types of cancer chemotherapy. Genetic selection in tumors and the consequent survival of multidrug resistant (MDR) cells is thought to account for this phenomenon. Endicott and Ling, 1989; Gottesman and Pastan, 1988. Multidrug resistance was originally characterized in vitro after the selection of actinomycin D or colchicine resistant cells. Bech-Hansen et al., 1976; Biedler and Riehm, 1970. These cells were shown to be resistant to a wide range of other drugs as well, such as adriamycin, vinblastine and puromycin.

The drugs to which these cells are resistant are dissimilar in both structure and mode of action. The only common features are that the drugs are hydrophobic, amphipathic compounds generally derived from natural products. Ling and co- workers showed that this multidrug resistance was correlated with the presence of a 150-170,000 molecular weight glycoprotein, which they termed P-glycoprotein. Riordan and Ling, 1985.

It is thought that the P-glycoprotein forms a molecular pump, which actively causes the excretion or efflux of drugs from the cell membrane or cytoplasm.

Endicott and Ling, 1989; Gottesman and Pastan, 1988; Horio et al., 1988. While the P-glycoprotein was originally studied in rodent cells, elevated levels have now been found in human cells that have been selected for drug resistance in vitro, as well as in multidrug resistant cancers. Goldstein et al., 1989. While increased amounts of P-glycoprotein appear to be important for the development of some cases of multidrug resistance, it does not appear to be the only mechanism by which cells can become multidrug resistant. Several workers have reported that cells selected for multidrug resistance show evidence of non-P- glycoprotein mediated multidrug resistance mechanisms, Beck et al., 1987; Cole et al., 1991; Shen et al., 1991, and multidrug resistant cells may exhibit more than one resistance mechanism simultaneously. Zijlstra et al., 1987.

There is also accumulating evidence that protein kinases may be involved in the expression and function of the P-glycoprotein and multidrug resistance. Several studies have described an increase in kinase activities or increased phosphorylation of the P-glycoprotein in multidrug resistant cells. Carlsen et al., 1977; Center, 1983; Center, 1985; Fine et al., 1988; Hamada et al., 1987; Mellado and Horwitz, 1987; Posada et al., 1989. Higher levels of protein kinase C are associated with increased multidrug resistance of some but not all multidrug resistant cells. Chambers et al., 1990; Fine et al, 1988; Posada et al., 1989. Yu et al. [1991] have shown that transfection of a plasmid expressing functional protein kinase Cα is associated with an increase in multidrug resistance. The inventors of this invention have shown that cAMP-dependent protein kinase levels are directly associated with relative multidrug resistance, Abraham et al., 1987, and P-glycoprotein mRNA and protein levels in Chinese hamster ovary cells. Abraham et al., 1990. These studies suggest that protein kinases may have a role in modulating multidrug resistance.

Full citations of the references herein are provided towards the end of the application.

We now describe the isolation, purification and structure of a 170 kDa membrane associated protein kinase. Summary of the Invention

This invention comprises a gene, gene fragment or cDNA coding for any member of a protein kinase family comprising DNA having a partial DNA sequence consisting of the cDNA sequence shown in Table 1, or the sequence given by Sequence ID no. 1, or an equivalent sequence, or a similar sequence having conservatively modified variations to the DNA sequences described by the DNA sequence shown in Table 1, or the sequence given by Sequence ID no. 1, or the cDNA similar to the DNA of the DNA sequence shown in Table 1, or the sequence given by Sequence ID no. 1, except that the cDNA similar to the DNA shown in Table 1, or the sequence given by Sequence ID no. 1, may contain codons coding for any of the amino acids in Table 3, shown as Yakl, Cdk4, PKA, PKC amino acids, that appear in the same column and same position as the amino acid sequence of PKR, but are different than the amino acids of PKR, these codons being substitutes or additions to the codons of the DNA sequence shown in Table 1, such substitutions or additions beginning with the codon that begins at position 535 (TAT) and following in the order shown in Table 3, that allow substitutions or additions to the cDNA sequence in Table 1, or the sequence given by Sequence ID no. 1, that code for amino acids in the same order as are provided by the PKR sequence of Table 3 only including the chosen additions or substitutions.

Described herein are genes, gene fragments or cDNA, that code for a membrane associated protein kinase belonging to this family of protein kinases, or having similar or identical gene sequences as those of Table 1 or sequence ID no. 1, and having a molecular weight of about 170 kD.

Described herein are genes, gene fragments or cDNA that coding for any member of a protein kinase family comprising a DNA sequence that codes for the peptide sequence shown in Table 1, or the sequence given by Sequence ID no. 4, or an equivalent sequence, or a similar sequence having conservatively modified variations to the DNA sequences that would code for the sequences consisting of the peptide sequence shown in Table 1, or the sequence given by Sequence ID no. 4, or an equivalent sequence, or a similar sequence having conservatively modified variations to the DNA sequences described by the peptide sequence shown in Table 1, or the sequence of Sequence ID no. 4, or the cDNA coding for the peptide sequence shown in Table 1, or the sequence given by Sequence ID no. 4, except that the cDNA coding for the peptide similar to the peptides shown in Table 1, or the sequence of Sequence ID no. 4, may contain codons coding for any of the amino acids in Table 3, shown as Yakl, Cdk4, PKA, PKC amino acids, that appear in the same column and same position as the amino acid sequence of PKR, but are different than the amino acids of PKR, these codons being substitutes or additions to the codons of the DNA sequence shown in Table 1, such substitutions or additions beginning with the codon that begins at position 535 (TAT) and following in the order shown in Table 3, that allow substitutions or additions to the cDNA sequence in Table 1, or the sequence given by Sequence ID no. 1, that code for amino acids in the same order as are provided by the PKR sequence of Table 3 only including the chosen additions or substitutions.

Described herein are any genes, gene fragments or cDNA, that codes for a membrane associated protein kinase belonging to this family of protein kinases, or having similar or identical gene sequences as those of Table 1 or sequence ID no. 1, and having a molecular weight of about 170 kD.

Described herein are any genes, gene fragments or cDNA coding for any member of a protein kinase family comprising a DNA sequence that codes for the peptide sequence shown in Table 1,

Described herein are any genes, gene fragments or cDNA, that codes for a membrane associated protein kinase belonging to this family of protein kinases, or having similar or identical DNA sequences that codes for the peptide sequences shown in Table 1, or sequence ID no. 4 or having substitutions as described above, and having a molecular weight of about 170 kD.

Described herein are peptides or proteins comprising the peptides having the amino acid sequence shown in Table 1 or shown by Sequence I.D. no. 4, or an equivalent peptide, or a similar peptide having conservatively modified variations, or a peptide comprising the peptide shown in Table 1, or by Sequence ID no. 4 except the equivalent peptide contains any 1 to 243, or any 1 to 20 of the amino acids in Table 3 shown as belonging to Yakl, Cdk4, PKA, or PKC that appear in the same column and same position as the amino acid sequence of PKR, but are different than the amino acids of PKR, these amino acid additions or substitutions begin at position 179 (Tyr) and follow in the same order as shown by Table 3, to make from any 1 to 243, or any 1 to 20, substitutions or additions to the peptide of Table 1 or Sequence ID no. 4.

Described herein are any peptides or proteins for a membrane associated protein kinase belonging to this family of protein kinases, or having amino acid substitutions or additions as described above, and having a molecular weight of about 170 kD.

Described herein are genes, gene fragments or cDNA coding for any member of a protein kinase family comprising the sequential amino acids in Table 3, shown as Yakl, Cdk4, PKA, PKC amino acids, that appear in the same column and same position as the amino acid sequence of PKR, but are different than the amino acids of PKR, these codons being substitutes or additions to the codons of the DNA sequence shown in Table 1, such substitutions or additions beginning with the codon that begins at position 535 (TAT) and following in the order shown in Table 3, that allow substitutions or additions to the cDNA sequence in of PKR, Table 3.

Described herein are peptides or proteins comprising the peptides having the amino acid sequence shown in Table 3, shown as PKR, or the amino acid sequence shown in Table 3, that appear in the same column and same position as the amino acid sequence of PKR, but are different than the amino acids of PKR, and that are taken from the Yakl, Cdk4, PKA, PKC amino acids, that appear in the same column and same position as the amino acid sequence of PKR, such substitutions or additions beginning with the first amino acid shown in Table 3, including all possible substitutions or additions to the cDNA sequence in of PKR, Table 3, from all the shown sequences, and continuing to the last amino acid shown in Table 3, including PKR, Yakl, Cdk4, PKA, PKC. Additional Description of the Invention and Description of the Preferred Embodiment(s)

Described here in detail is a novel kinase, a 170-kDa serine-threonine kinase designated 170 kDa kinase, or "PKR," and the gene that codes for this kinase, "PKR." This so called PKR gene and protein is probably more aptly named PKY, because the term "PKR" has recently been used to refer to a different kinase, an interferon induced, double stranded, RNA activiated protein kinase, see Polyak S.J., Tang N., Wambach M., Barber G.N., and Katze M.G., Journal of Biological Chemistry, 271(3): 1702-1707, 19 January 1996, "The P58 Cellular Inhibitor Complexes with the Interferon-Induced, Double-Stranded RNA-Dependent Protein Kinase, PKR, To Regulate Its Autophosphorylation and Activity." However, to maintain consistency between this application and priority applications, the PKR term is used herein. In this document, the letters PKR may refer to the gene, or cDNA, the full protein, or peptides of the full protein, depending on the context in which the abbreviation is used. Definitions Some abbreviations used are: MDR, multidrug resistant; DME, Dulbecco's modified Eagle's medium; FBS, fetal bovine serum; SDS, sodium dodecyl sulfate; PVDF, polyvinylidene difluoride; BSA, bovine serum albumin; EGF, epidermal growth factor; PBS, Dulbecco's phosphate-buffered saline; TN, 10 mM Tris-HCl, pH 7.4, 0.5 M NaCl; PMA, phorbol 12-myristate 13-acetate. The sequences included with this disclosure are provided as Sequence Identification Numbers 1-13, and listed toward the end of this document. These sequences are also called, sequence ID numbers or sometimes, sequence numbers, "bp" means base pair or base pairs. Amino acids described herein are abbreviated with typical one or three letter codes. The sequence Usting, produced with the Patent In© program, uses 3 letter codes for amino acids.

Compounds of the Invention

Identification and Isolation

The C terminal portion of the gene that codes for the kinase, PKR, is described by Table 1 and sequence number 1, and the C terminal portion of the novel kinase, a 170-kDa serine-threonine kinase designated 170 kDa kinase, is provided in sequence number 4. The asterisk (@) in Table 1, and Sequence I.D Number 1, indicates an unspecified amino acid occurs in that position. The sequences in Table 1 and the sequence I.D. numbered listings begin with the portion of the N-terminal sequence that is known. We believe the known protion represents the C-terminal portion of the DNA and the corresponding protein.

A PCR (polymerase chain reaction) based method was used to screen for the gene, PKR, that encodes 170 kDa kinase. Woodgett et al., 1991; Schultz et al., 1993. cDNA was made from mRNA isolated from the multidrug resistant human cell line, KB-V1. Specific oligonucleotides pairs, whose sequence was based on short conserved regions of serine-threonine kinases were used as templates to hybridize to KB-V1 cDNA in a PCR reaction. cDNA sequences that were homologous to conserved regions of serine-threonine kinases were amplified. Hanks et al., 1989. The 170 kDa kinase gene (PKR) was recovered using primers spanning kinase homology domains VI and DC. Using these primers, 150-220 bp fragments were predicted based on the conserved spacing of kinase homology domains. Hanks, et al 1988. Several of the fragments within the predicted class were synthesized and a 193 bp fragment was PCR amplified and recovered.

Putative kinase fragments were identified by the presence of the consensus DFG amino acid sequence at the proper position in the predicted amino acd sequence. An aspartic acid(D) at 12-18 amino acids 3' to the DLKPEN sequence (beginning at amino acid 525 in PKR) is invariant in all kinases. The residues surrounding aspartic acid (D), phenylalanine (F) and glycine (G) are commonly conserved and are present in PKR. The identity of the PCR fragments was further identified by hybridization to mRNA in Northern blots. One of these PCR fragments, a 193 bp fragment, detects a message that is overexpressed in four different multidrug resistant cell line as compared to the sensitive parental line and detects a message of 7 kb. The amplified and isolated PCR fragments were used to recover larger cDNA clones from a cDNA library made from a multidrug resistant cell, KB-V1. After sequencing, these clones predict a protein of over 1403 amino acids that has a high degree of homology to many serine-protein kinases. Its highest homology is to the yeast Yakl protein kinase in yeast and the CDK (cell cycle dependent kinases ) family of protein kinases in humans. The 170 kDa kinase is membrane-associated and has optimal activity in

10 mM Mn⁺⁺.

Previous reports have shown that the EGF receptor is elevated in some multidrug resistant cells (Meyers et al., 1986; 1988). Since the EGF receptor is similar in size to 170 kDa kinase, we investigated whether it was identical to the 170 kDa kinase. The 170 kDa kinase was established to be smaller than the EGF receptor in A431 and KB cells after comparing the molecular weights of the 170 kDa kinase identified on the in situ activity blots and the EGF receptor identified either by anti-phosphotyrosine Westerns or immunoprecipitation with an anti-EGF receptor antibody. The fact that the 170 kDa kinase was not recognized by anti-phosphotyrosine monoclonal antibody suggests that it is not a tyrosine kinase. Other experiments showed that the radiolabelled phosphate residue(s) of the 170 kDa band is easily removed by prolonged KOH treatment, also suggesting that it is not a tyrosine kinase. In addition, phosphoamino acid analysis of proteins eluted from the PVDF membrane in the area of the 170 kDa kinase did not show any evidence of phosphotyrosine. The presence of phosphothreonine and some phosphoserine in the phosphoamino acid digests suggest instead that the 170 kDa kinase is a serine- threonine kinase. This result also supports the evidence that the 170 kDa kinase is not P-glycoprotein, since P-glycoprotein appears to be phosphorylated mainly on serine residues (Hamada et al., 1987).

This kinase probably has structural similarities to other known serine- threonine kinases since it is inhibited by the serine/threonine kinase inhibitors staurosporine, K252a, and KT5720. However, the large size of this kinase makes it unusual, since most identified serine/thronine kinases are less than 100 kDa in size. In a study using the in situ kinase assay, Ferrell and Martin (1989) identified a 170 kDa kinase (PK170) in whole cell lysates of human platelets and in mouse NIH 3T3 cells (Ferrell and Martin 1990). Activity of the platelet kinase as demonstrated in the in situ assay was increased when platelets were pretreated in vivo with thrombin or both phorbol 12-myristate 13-acetate (PMA) and ionomycin. They postulated that this increase in activity might be due to a change in the phosphorylation state of the kinase. The 170 kDa kinase described herein shows no stimulation when KB-V-1 cells are pretreated with PMA or ionomycin, in combination or separately (not shown).

The 170 kDa kinase described herein thus appears to be distinct from the platelet-derived 170 kDa kinase. However, signalling mechanisms present in the platelet cell that affect 170 kDa kinase may not be present in the KB-Vl cells that were investigated here. Interestingly, the 170 kDa kinase described by Ferrell and Martin, and the 170 kDa kinase described herein, both appear to cause phosphorylation principally on threonine. The 170 kDa activity of the protein described herein was found associated almost exclusively with the membranes, unlike the Ferrell and Martin enzyme.

The large size and membrane localization of the 170 kDa kinase described herein suggests that it may have some type of receptor function. A new class of membrane-bound receptors with serine-threonine kinase activity that recognizes ligands in the TGF family has recently been identified [Massaque, 1992]. This kinase may play a similar role.

Staats et al. [1990] described a membrane associated phospholipid-stimulated kinase, found in both drug sensitive HL60 and in multidrug resistant HL60/Vinc, cells which are capable of phosphorylating the P-glycoprotein in vitro. The 170 kDa kinase activity described here, however, was found only in the drug resistant KB-Vl cells and not in the parental sensitive KB-3-1 cells, although it was found in some other sensitive cells. The 170 kDa kinase studied here showed optimal activity with Mn⁺⁺, similar to the enzyme described by Staats et al. , but unlike that enzyme, the enzyme described herein is not stimulated by phosphatidylserine. It is possible that activation by phosphatidylserine might not occur in the in situ assay since kinases were presumably no longer associated with membrane; however, membrane fractions of HL60 cells used in the in situ assay showed no 170 kDa kinase activity (data not shown). Staats et al. suggested that the HL60 kinase was similar to protein kinase P, a platelet kinase that is described by Elias and Davis (1985) as 27 kDa. These results suggest that the 170 kDa kinase is different from the described HL60 kinase.

Because P-glycoprotein has been reported to have a molecular weight of 170 kDa in KB-Vl cells, we performed experiments to determine if it was a distinct molecule from the 170 kDa kinase described herein. All the tested multidrug resistant cells were shown to express P-glycoprotein by Western blot analysis. The molecular weight of the P-glycoprotein varied from 150 to 180 kDa in these cell lines, possibly due to differences in glycosylation. In comparison, the 170 kDa kinase molecular weight was invariant. Because the 170 kDa kinase activity was seen in some cells which do not express the P-glycoprotein, and because of the difference in molecular weight of the P-glycoprotein in some cell lines, we conclude that the 170 kDa kinase is not the same protein as P-glycoprotein.

Cells transfected with the MDR1 gene and selected for vinblastine resistance did express the P-glycoprotein, but did not have 170 kDa kinase activity. The lack of the 170 kDa kinase in the transfected cells definitively shows that this kinase activity is not due to P-glycoprotein.

The kinase is also widely distributed. It has been found in the rat brain in addition to the cells described above. The characteristics of the rat brain membrane 170 kDa protein kinase are identical to that of the 170 kDa kinase from MDR cells. The autophosphorylated 170 kDa kinase from rat brain has the identical phosphoamino acid composition and phosphopeptide map after EndoLys C digestion as that isolated from MDR KB-Vl cells. RNA coding for the 170 kDa kinase, described herein (PKR), has also been found in high abundance in heart and muscle tissue. This finding suggests the protein may play an important role in ion transport or regulation. Sequence of the PKR Clone

Screening of a KB-V-1 cDNA library with the PCR PKR fragment yielded a single 3.5 kb cDNA clone. The 3.5 kb clone was isolated using the 193 bp PCR fragment, described above, as a probe. Two cDNA clones with flanking regions were recovered using a 900 bp 3' Pstl fragment and a 600 bp 5' Pstl fragment as probes. A large and significant portion of the cDNA has been sequenced and is claimed herein. 3605 bps of sequence have been obtained from cDNA PKR and this sequence spans an open reading frame that predicts a 1197 amino acid protein. In addition to this precisely identified peptide we believe there are an additional 350 or so amino acids coded for by about 1050 bps that are not yet precisely identified. The predicted amino acid sequence of the known peptide sequence based upon the 3605 described bps is somewhat homologous to many protein kinases. The predicted amino acid sequence of the precisely identified portion of the protein spans the conserved 280 amino acid catalytic core of serine-threonine protein kinases and contains all 11 invariant amino acids in the appropriate positions. Further, the lysine residue at amino acid 182 is indicative of a serine/threonine kinase and is never found in a tyrosine kinase.

The described sequence is indicative of a serine/threonine kinase but has certain unique features. PKR has a larger than normal distance between homology domains DC and XI. Most kinases have about 40 amino acids separating these domains, PKR has 75. Only Yak-1 and CDC7 have a similar structure. Yak-1 has in particular a rather high level of homology to PKR. In the catalytic core region, the Yak-1 amino acid sequence is 50% identical to PKR. Eighty percent (80%) of the amino acids of Yak-1 (core region) show similarity to PKR. Similarity refers to the sum of amino acids that are identical or conservatively changed for a particular region. The changes should not affect the charge or size. This high level of similarity contrasts with the core regions of CDK-4, PKC, and PKA sequences which have between 25-30% identity and 50-67% similarity. The homology to Yak-1 is also much longer, extending to the end of the Yak-1 sequence. All homology to known human kinases ends after the catalytic core sequences. The sequences shown in Table 1 are based on the sequence of a cDNA clone derived from reverse transcription of mRNA. The asterisk (@) in Table 1, and Sequence I.D Number 1, indicates an unspecified amino acid occurs in that position. Sequencing of a genomic DNA fragment covering this gene suggests that there is some minor polymorphism in the sequence and that some conservative changes in the sequence may occur in different individuals. In general, minor polymorphism do not alter the amino acid sequence; however, even minor changes in the amino acid structure could be tolerated, provided they do not significantly alter the activity of the enzyme, see Table 3 and below.

PKR thus is a newly described mammalian kinase that is highly homologous to the Yak-1 protein in yeast. It is the first member of what we believe will be a class of similar human kinases that are somewhat homologous to Yakl.

A portion of the amino acid sequence of the 170 kDa kinase coded for by PKR is compared to the sequence for Yakl, Cdk4, PKA and PKC in Table 3. This table shows that many amino acid substitutions could be made in the PKR gene. The invention discloses, describes and claims all such variations as may be suggested by Table 3, as novel gene and protein or peptide sequences. One of ordinary skill in the art would be able to use Table 3 to create homologous variants of the PKR gene that retained activity. All homologous variants suggested by Table 3, variants suggested by other known sequences of similar kinases, variants obvious to one of ordinary skill in the art are hereby disclosed, described and claimed. The sequences shown in Table 3 correspond to portions of PKR, Yakl, Cdk4, PKA and PKC. The entire known cDNA sequence of PKR is shown in Table 1 and the sequence description for sequence ID No. 1. The asterisk (@) in Table 1, and Sequence I.D Number 1, indicates an unspecified amino acid occurs in that position. The sequences shown in Table 3, showing PKR sequences, begin at a sequence that begins after counting down 534 bases from the beginning of sequence ID No. 1. That is, only a portion of the full PKR, Yakl, Cdk4, PKA and PKC sequences are shown in Table 3. Table 3 is prepared so that corresponding amino acids for Yakl, Cdk4, PKA and PKC can be compared to the amino acids at corresponding positions on the PKR protein. Different amino acids sequences composed of Yakl, Cdk4, PKA and PKC amino acids corresponding to the same position as amino acids from PKR should be obvious and are claimed. An asterisk (*) shows that at some positions on PKR (marked with an asterisk "*") there could be an amino acid that is equivalent to the corresponding amino acid in the corresponding peptides (Yakl, Cdk4, PKA and PKC). In this situation an appropriate and corresponding protein similar to PKR would be a PKR having an extra amino acid at the position of the asterisk (*).

The following examples of homologous sequences are provided. Research described herein and the scientific knowledge of the inventors indicates that the following types of homologous gene and protein sequences should produce proteins having similar activity to the peptide/protein described by Sequence Id. No. 4 for genes and proteins having: Genes that are >80 % homologous or have >80 % conservative changes in the core region and >40 % homologous or have >40 % conservative changes in the noncore regions. Genes that are >80 % homologous or have >80 % conservative changes in the core region and >50 % homologous or have >50 % conservative changes in the noncore regions. Genes that are >90 % homologous or have >90 % conservative changes in the core region and >60 % homologous or have >60 % conservative changes in the noncore regions. Genes that are >90 % homologous or have >90 % conservative changes in the core region and >40 % homologous or have >40 % conservative changes in the noncore regions. Genes that are >90 % homologous or have >90 % conservative changes in the core region and >50 % homologous or have >50 % conservative changes in the noncore regions. Genes that are >90 % homologous or have >90 % conservative changes in the core region and >60 % homologous or have >60 % conservative changes in the noncore regions. An isolated gene having conservatively modified variations of the DNA sequence described by Sequence Identification Number 1 or Table 1. Conservatively modified variations, or conservatively modified changes means that amino acids with similar charges or side chains may be substituted for the original amino acids. These changes should not substantially affect the activity. The conservative changes should not substantially affect the hydrophobicity of the new variant. Even nonconservative changes would be acceptable outside of the conserved kinase region, see Hanks et al. 1988, incorporated by reference, for a discussion of the core region. Description o the Sequence Listings

Sequence listings are included with this document. Sequence number 1 is the precisely identified portion of the cDNA sequence of PKR that includes the known coding region. Sequence number 12 is the cDNA sequence shown in Table 1. Sequence number 2 is the precisely identified portion of the cDNA sequence of the portion of PKR that begins with the DNA coding for the conserved amino acids shown in Table 3 and ends with the same sequences of sequence number 1. Sequence number 3 is the precisely identified portion of the cDNA sequence of the portion of PKR that begins with the DNA coding for the first amino acid shown in Table 3 and ends with the DNA coding for the last amino acid shown in Table 3. Sequence number 4 is the amino acid sequence of the protein or peptide that is transcribed from the DNA sequence of sequence number 1. Sequence number 13 is the amino acid sequence of the protein or peptide shown in Table 1.

Sequence number 5 is the amino acid sequence of the protein or peptide that is transcribed from the DNA of sequence number 2. Sequences corresponding to sequence numbers 1, 2, 3, 4 and 5 can be found in Table 1. Sequence number 6 is the amino acid sequence of the highly conserved region of the protein that is transcribed from the DNA of sequence number 3. Sequence number β is in Table 3. Sequence number 7, also in Table 3, corresponds to sequence number 6 only additional possibilities for extra amino acids are allowed, the asterisk (*) from Table 3 is replaced with an X in the sequence listing. In one embodiment of this invention the X may, optionally, be substitued with any conservatively modified amino acid. In another embodiment of this invention the X may be substituted with any other amino acid from either Yakl, Cdk4, PKA and or PKC as indicated in Table 3, that is in the same position as the asterisk (*) is in. Sequence number 8 corresponds to the conserved peptide region of Yakl shown in Table 3. Sequence number 9 corresponds to the conserved peptide region of Cdk4 shown in Table 3. Sequence number 10 corresponds to the conserved peptide region of PKA shown in Table 3. Sequence number 11 corresponds to the conserved peptide region of PKC shown in Table 3.

Examples of substitutions. Consider the first peptide, "PKR," in Table 3. This peptide is also provided in sequence number 6. The first amino acid shown for the peptide "PKR" in Table 3 is Y or Tyrosine. Tyrosine is also the first amino acid shown for the peptides Yakl and PKA; however, the first amino acid shown for the peptide PKC is N, or Asparagine. Thus, a peptide that is identical to the PKR peptide in Table 3 is shown and described and so is a peptide being only one amino acid different than the PKR peptide shown in Table 3. This other peptide and protein is PKR plus, having asparagine not tyrosine at this position. The second position of the PKR peptide, from Table 3, is Q or Glutamine, in addition, Table 3 shows that the second position can also be I (isoleucine), R (arginine) or K (lysine), or any substitution thereof. This disclosure thus describes all combinations possible for the peptides and proteins of PKR and the possible amino acids shown in Table 3. Especially preferred are combinations where from 1 to 20 substitutions or additions are made. So, the first position may be argaragine and the second position isoleucine. All combinations and substitutions shown in Table 3 are described for the full protein, and peptides having the conserved region. That is, the peptides shown in Table 3, labeled "PKR," and similar amino acid substitutions or additions may be made to the full precisely identified peptide shown in Table 1, or as sequence number 4. The sequence shown in Table 3 is a part of sequence number 4.

Note that the "*" in Table 3 indicates that there is no amino acid in that position for that particular peptide; however, there could be an amino acid substitution made in that position, as Table 3 indicates. Thus, the sequence of PKR at the peptide postion shown in Table 3 for positions 9, 10 and 11 is as follows: For PKR the sequence is ... L*C ... (Leucine - Cysteine) but the same sequence for Yak 1 is ... LGV ... (Leucine-Glycine-Valine). The asterisk (*) in the PKR sequence, in Table 3, indicates that PKR normally has no amino acid at the place where the Yak 1 gene has a Glycine. (Also note that here the cysteine of position 11 could be replaced with valine, threonine or lysine.)

Table 3 shows that each amino acid in the conserved region could be assigned a position. Position one (1) of these peptide segments for PKR, Yakl and PKA is occupied by tyrosine (Y). PKC has a position 1 occupied with asparagine (N). Herein is described then, proteins and peptides identical to either the full length PKR or smaller segments, such as the one shown in Table 3 where different amino acids in the same position could be substituted for the original PKR amino acids. Position 1 is the position of the first amino acid shown in Table 3. (Position 1 from Table 3 and sequence ID no. 6, 7, 8, 9, 10, 11 corresponds to amino acid position 179 of Table 1. Position 1 from Table 3 also corresponds to the codon TAT that begins at position 535 of the DNA sequence of Table 1.) The first asterisk (*) in the PKR sequence, Table 3, indicates that PKR normally has no amino acid at the place where the Yak 1 protein has a glydne. Therefore, Table 3 should be interpreted as showing that PKR can have a glycine in this position, in addition to the other PKR amino acids. This position, of the first asterisk (*), position 10, would correspond to the place between amino acids 187 and 188 (Leu and Cys) of the amino adds in Table 1 and sequence ID 3. Table 3 also shows that cysteine at postion 11 could be substituted with valine, threonine or lysine at position 11. Positions in the peptide sequence shown in Table 3 do not always make one to one correspondent matches with amino adds from Table 1 or sequence no. 1, because extra amino acids are possible, as shown by an "*" in Table 3. All possible combinations and substitutions shown in Table 3 are described. All possible corresponding combinations and corresponding substitutions shown in Table 3 are also applicable and described for the full length protein or peptide shown in Table 1, sequence ID numbers 4-6, as well as the corresponding DNA, sequence numbers 1-3. Sequence number 6 shows an X where Table 1 shows a "*," but note that substitutions to the sequence are not limited to sequences having an X, the X only shows positions where additional amino adds not normally found in PKR may be added. Substitutions may be made any place they are suggested by alternate amino adds shown in Table 3 or any other part of this document. Minor polymorphisms and conservative changes may occur at any point in the DNA sequence and minor changes not significantly affecting the activity of the enzyme may occur in the peptides or protein. Elevated levels of mRNA The lβ BlβAdr¹¹, HLβO/fflβOAdr¹¹, MCF7/MCF7Adr^R, and

PSδS/PSSSVin^SSβAdr¹¹ cell lines were used to determine if the message detected by PCR fragments were overexpressed in other resistant cell lines in addition to KB- V-l. PKR was expressed at higher levels in several multidrug resistant cell lines as compared to their parental sensitive cells. It had higher expression in the HLβO/Adr¹* ,P388Vin^R, and KB-Vl cell lines as compared to the sensitive parental lines, MCF7, HL60, P388 and KB-3-1, respectively. See Table 2. MCF7Adr^R resistant cells also expressed PKR at higher levels than the parental sensitive cells, MCF7. However, the multidrug resistant cell line BlβAdr¹¹ expressed slightly lower levels in the resistant cell line as compared to the sensitive cells. See Table 2. This lack of significant increase in expression in the BlβAdr^ resistant cells could be explained by the relatively high level of 170 kDa kinase (PKR) expression in the B16 sensitive cells. Relative levels of expression of PKR protein and mRNA expression are presented in Table 2 and show that there is a good correlation between level of 170 kDa kinase (PKR) kinase activity and mRNA levels. Utility of the Invention

The novel kinase, 170 kDa kinase, has higher activity in several multidrug resistant cell lines compared to their sensitive parental cells. It is also expressed at high activity in the brain. The 170 kDa kinase may be very important in the pathway leading to multidrug resistance, and inhibition of this kinase may lead to a reversal of drug resistance. This would have therapeutic importance in re- sensitizing drug resistant cancer cells to the effects of chemotherapeutics. PKR, the isolated gene, codes for the 170 kDa kinase. The protein's large size and membrane localization suggests that it may have some receptor function.

In addition to its role in multidrug resistance, the inventors have discovered evidence of significant levels of the protein in heart, muscle and brain tissue. The discovery of significant levels of the protein in these organs suggests the protein has important functions.

A somewhat homologous protein is found in yeast. Yakl, also codes for a protein kinase. The Yakl protein of yeast has a 35% identical amino acid sequence over 450 amino adds. After Yakl, the kinase that is most similar to PKR is the CDK (cyclin dependent kinase) family that has an average of 30% identical amino acid sequence over 285 amino acids. The high level of homology to Yakl may provide some indication of the function of PKR. Yakl has been shown to be involved in the cell-cycle regulatory pathway. Yakl acts to inhibit cell growth in yeast in a pathway that is parallel and antagonistic to that of PKA(cAMP-dependent protein kinase) (Garrett et al., 1991).

Overexpression of Yakl induces a thermotolerant phenotype (Hartley et al., 1993). In analogy to the effect of Yakl, overexpression of the PKR gene may have cytoprotective effects; cells with an activated stress-tolerance pathway may be at a selective advantage. Cells that have altered their metabolism by becoming multidrug resistant through over-expression of P-glycoprotein, MRP, or other mechanisms, or as a response to drug treatment itself may elicit such a stress response. If over-expression of the PKR is a stress response then inhibition of this kinase will lead to toxicity of multidrug resistant cells. Inhibition of this enzyme will therefore be expected to re-sensitize cells to chemotherapeutic drugs. Should PKR, and its protein, have a role in heat resistance or stress control response, then inhibition might lead to death of resistant tumor cells. Activation or increased synthesis of this gene may have a protective effect in a variety of cells. Full References Provided Abraham ., Chin, K-V, Gottesman, M.M., Mayo, J.K.Sampson, KE. (1990)

Transfection of mutant regulatory subunit gene of cAMP-dependent protein kinase causes increased drug of a 170 kDa membrane kinase with increased activity in KB- Vl multidrug resistant cells. J. Cell. Biochemistry 52:384-395.

Abraham, I., Hunter, R.J., Sampson, KE., Gottesman, M.M., and Mayo, J.K. 1987. Cyclic AMP-dependent protein kinase regulates sensitivity of cells to multiple drugs. Mol. Cell. Biol. 7:3098-3106.

Abraham, I., Chin, K-V., Gottesman, M.M., Mayo, J.K, and Sampson, KE. 1990. Transfection of a mutant regulatory subunit gene of cAMP-dependent protein kinase causes increased drug sensitivity and decreased expression of the P- glycoprotein. Exp. Cell. Res. 189:133-141.

Balakrishnan, R., Frohlich, M.W., Rahaim, P.T., Backman, K, and Yocum, R.R. (1993). Cloning and sequence of the gene encoding enzyme E-l of the methionine salvage pathway of Klebsiella Oxytoca. J. Biol. Chem. 268:24792-24795. Begley and Abraham (1994) Protein kinase sequences expressed in multidrug resistant KB-Vl cells. FASEB j. 8:A1392.

Chambers, T.C. McAvoy, E.M., Jacobs, E.W., and Eilon, G. 1990. Protein Kinase C phosphorylates P-glycoprotein in multidrug resistant human KB carcinoma cells. J. Biol. Chem. 265:7679-7686.

Chou, P.Y., Fasman, G.O. (1978) Prediction of the secondary structure of proteins from their amino acid sequence. Adv. Enz. 47:45-147.

Coffer, J., and Woodgett, J.R. 1991. Molecular cloning and characterization of a novel putative protein-serine threonine kinase related to the cAMP-dependent and protein kinase C families. J. Biochem. 201:475-481.

Cole,S.P., Bhardwaj.G., Gerlach,J.H., Mackie,J.E., Grant.C.E., Almquist,KC, StewartA ., Kurz,E.U., Duncan -M*. Deeley,R.G.1992 Overexpression of a transporter gene in a multidrug-resistant human lung cancer cell line. Science 258: 1650-.

Elias, L., and Davis, A. Manganese-phospholipid-stimulated protein kinase activity of human leukemic cells. J. Biol. Chem., 260:7023-7028, 1985. Feinberg, A.P., Vogelstein, B. (1984 ) A technique for radiolabeling DNA restriction endonudease fragments to high specific activity. Addendum Anal-Biochem. 137:266-7.

Ferrell, J.E., Jr., and Martin G.S. Thrombin stimulates the activities on multiple previously unidentified protein kinases in platelets. J. Biol. Chem., 264:20723-20729, 1989.

Ferrell, J.E., Jr., and Martin, G.S. Identification of a 42-kilodalton phosphotyrosyl protein as a serine (threonine) protein kinase by renaturation. Mol. Cell. Biol., 20:3020-3026, 1990.

Fine, R.L., Patel, J., and Chabner, B.A. 1988. Phorbol esters induce multidrug resistance in human breast cancer cells. Proc. Nat. Acad. of Sci. USA 85:582-586.

Garrett, S., Menold, M.M., and Broach, J.R. 1991. The Saccharomyces cereυisiae Yakl gene encodes a protein kinase that is induced by arrest early in the cell cycle. Mol. and Cell. Biol. ll(8):4045-4052. Hanks, S.K., Quinn,A.M., Hunter,T ( 1988)The protein kinase family: conserved features and deduced phylogeny of the catalytic domains. Science 241: 42-52.

Hamada, H., Hagiwara, K-L, Nakajima, T., and Tsuruo, T. Phosphorylation of the M_j. 170,000 to 180,000 glycoprotein specific to multidrug-resistant tumor cells: effects of verapamil, trifluoperazine, and phorbol esters. Cancer Res., 47:2860-2865, 1987.

Hartley, A.D., Ward, M.P., and Garrett, S. 1994. The Yakl protein kinase of Saccharomyces cereυisiae moderates thermotolerance and inhibits growth by an Sch9 protein kinase-independent mechanism. Genetics 136:465-474. Massaque, J. Receptors for the TGF-β family. Cell, 69: 1067-1070, 1992.

Meyers, M.B., Merluzzi, V.J., Spengler, B.A., and Biedler, J.L. Epidermal growth factor receptor is increased in multidrug-resistant Chinese hamster and mouse tumor cells. Proc. Natl. Acad. Sci. USA, 83:5521-5525, 1986.

Meyers, M.B., Shen, W.P.V., Spengler, B.A., Ciccarone, V., O'Brien, J.P., Donner, D.B., Furth, M.E., and Biedler, J.L. Increased epidermal growth factor receptor in multidrug-resistant human neuroblastoma cells. J. Cell. Biochem., 38:87-97, 1988.

McCroskey, M.C., Sampson, KE., Abraham, I. Isolation and partial characterization of a 170 kDa rat brain membrane associated protein kinase. ASBMB/DBC-ACS Joint Federation Meeting, May 21-25, 1995, abstract.

Posada, J.A., McKeegan, E.M., Worthington, KG., Morin, M.J., Jaken, S., and Tritton, T.R. 1989. Human multidrug resistant KB cells overexpress protein kinase C: involvement in drug resistance. Cancer Commun. 1:285-292.

Sampson, KE., McCroskey, M.C. and Abraham, I. (1993) Identification of a 170 kDa membrane kinase with increased activity in KB-Vl multidrug resistant cells. J. Cell. Biochemistry 52:384-395.

Schulz, J.S., and Nigg, E.A. 1993. Identification of 21 novel human protein kinases, including 3 members of a family related to the cell cycle regulator nimA of Aspergillus nidulans. Cell Growth and Differentiation 4:821-830. Staats, J., Marquardt, D., and Center, M.S. Characterization of a membrane- associated protein kinase of multidrug-resistant HL60 cells which phosphorylates P- glycoprotein. J. Biol. Chem., 265:4084-4090, 1990.

Riordan, J.R., and Ling, V. 1985. Genetic and Biochemical characterization of multidrug resistance. Pharmacol. Ther. 28:51-75. Yu, G., Ahmad, S., Aquino, A, Fairchild, C.R., Trepel, J.B., Ohno, S., Suzuki,

K, Tsuruo, T., Cowan, KH., and Glazer, R.I. 1991. Transfection with protein kinase C-alpha confers increased multidrug resistance to MCF-7 cells expressing P- glycoprotein. Cancer Commun. 3:181-189.

Table 1

The following polymer, cDNA, and amino acid sequences are provided. Note several sequences are provided, the entire sequence, the sequence begining at position 1 and extending to position 535, the sequence beginning at position 535 and extending to the end. The sequence beginning at position 535 and ending at the end of the codon that codes for the last amino acid in Table 3, and all combinations thereof. Corresponding segments, polymers, peptides or proteins composed of the amino acids described below, which correspond to the DNA seqments and are coded by those DNA segments are also provided. Polymorphisms are also included and other standard, routine, or obvious substitutions in the DNA or the amino acid polymers. The sequences provided correspond to the c- terminal end of the amino acid sequence. The true and absolute N- terminal end of the protein sequence and the corresponding DNA has not been completely identified.

CAAGTCTTGG TCTACCCACC ATATGTTTAT CAAACTCAGT CAAGTGCCTT TTGTAGTGTG 60

AAGAAACTCA AAGTAGAGCC AAGCAGTTGT GTATTCCAGG AAAGAAACTA TCCACGGACC 120

TATGTGAATG GTAGAAACTT TGGAAATTCT CATCCTCCCA CTAAGGGTAG TGCTTTTCAG 180 ACAAAGATAC CATTTAATAG ACCTCGAGGA CACAACTTTT CATTGCAGAC AAGTGCTGTT 240

GTTTTGAAAA ACACTGCAGG TGCTACAAAG GTCATAGCAG CTCAGGCACA GCAAGCTCAC 300

GTGCAGGCCC CTCAGATTGG GGTGTGGCGA AACAGATTGC ATTTCCTAGA AGGCCCCCAG 360

CGATGTGGAT TGAAGCGCAA GAGTGAGGAG TTGGATAATC ATAGCAGCGC AATGCAGATT 420

GTCGATGAAT TGTCCATACT TCCTGCAATG TTGCAAACCA ACATGGAAAA TCCAGTGACA 480 GTTGTGACAG CTACCACAGG ATCAAAACAG AATTGTACCA CTGGAGAAGG TGACTATCAG 540

*

TTAGTACAGC ATGAAGTCTT ATGCTCCATG AAAAATACTT ACGAAGTCCT TGATTTTGTT 600

GGTCGAGGCA CGTTTGGCCA GGTAGTTAAA TGCTGGAAAA GAGGGACAAA TGAAATTGTA 660

GCAATCAAAA TTTTGAAGAA TCATCCTTCT TATGCCCGTC AAGGTCAAAT AGAAGTGAGC 720

ATATTAGCAA GGCTCAGTAC TGAAAATGCT GATGAATATA ACTTTGTACG AGCTTATGAA 780 ACGTTTCAGC ACCGTAACCA TACTTGTTTA GTCTTTGAGA TGCTGGAACA AAACTTGTAT 840

GACTTTCTGA AACAAAATAA ATTTAGTCCC CTGCCTCTAA AAGTGATTCG GCCCATTCTT 900

CAACAAGTGG CCACTGCACT GAAAAAATTG AAAAGTCTTG GTTTAATTCA TGCTGATCTA 960

AAGCCTGAGA ATATTATGTT GGTGGATCCT GTTCGGCAGC CTTACAGGGT TAAAGTAATA 1020

GACTTTGGGT CGGCCAGTCA TGTATCAAAG ACTGTTTGGT CAACATATCT ACAATCTCGG 1080 TACAGAGCTC CAGAGATTAT ATTGGGGTTG CCATTTTGTA AGCCAATAGA CATGTGGTCA 1140

*Note, Table 3 starts with the aa (tyr) from the codon TAT beginning at 535. TTGGGATGTG TGATTGCAGA ATTATTTCTT GGATGGCCGC TATACCCAGG AGCCTTGGAG 1200

TATGATCAGA TTCGATACAT TTCTCAGACT CAAGGTTTGC CAGGAGAACA GTTGTTAAAT 1260 GTGGGTACTA AATCCACAAG ATTTTTTTGC AAAGAAACAG ATATGTCTCA TTCTGGTTGG 1320

AGATTAAAGA CATTGGAAGA GCATGAGGCA GAGACAGGAA TGAAGTCTAA AGAAGCCAGA 1380

AAATACATTT TCAACAGTCT GGATGATGTA GCGCATGTGA ACACAGTGAT GGATTTGGAA 1440

GGAAGTGATC TTTTTCCTGA GAAAGCTGAT AGAAGAGAAT TTGTTAGTCT GTTGAAGAAA 1500

ATGTTGCTGA TTGATGCAGA TTTAAGAATT ACTCCAGCTG AGACCCTGAA CCATCCTTTT 1560 GTTAATATGA AACATCTTCT AGATTTCCCT CATAGCAACC ATGTAAAGTC CTGTTTTCAT 1620

ATTATGGATA TTTGTAAGTC CCACCTAAAT TCATGTGACA CAAATAATCA CAACAAAACT 1680

TCACTTTTAA GACCAGTTGC TTCAAGCAGT ACTGCTACAC TGACTGCAAA TTTTACTAAA 1740

ATCGGAACAT TAAGAAGTCA GGCATTGACC ACATCTGCTC ATTCAGTTGT GGCGGATGGA 1800

ATACCTCTGC AGGCAGGAAC TCGTCAGTTT GGTTGTGGTG ATGCTTTTCA GCAGACATTG 1860 ATTATCTGTC CCCCAGCTAT TCAAGGTATT CCTGCAACAC ATGGTAAACC CACCAGTTAT 1920

TCAATAAGGG TACATAATAC AGTTCCACTT CTAACTCAGG CCCCAGCTGT GCAGCCACTA 1980

CAGATCCGAC CAAGGAGTTC TTTTCTCAGA CGTCCTCTGG TAGAACACAG ACAGATGCTG 2040

GTGCCTGCCT GGCAACAGGT GACACCCCTG GCTCCTGCTA CTACTACACT AACTTCTGAG 2100

AGTGTGGCTG GTTCACACAG GCTTGGAGAC TGGGGGAAGA TGATTTCATG CAGCAATCAT 2160 TATAACTCAG TGATGCCGCA GCCTCTTCTG ACCAATCAGA TAACTTTATC TGCCCCTCAG 2220

CCAGTTAGTG TGGGGATTGC ACATGTTGTC TGGCCTCAGC CTGCCACTAC CAAGAAAAAT 2280

AAACAGTGCC AGAACAGGAG TAATTCATTA CAGAATACCA ATATCCCACT ATCAGCATTT 2340

ATTTCTCCAA AGATAATTAA TGGGAAAGAT GTCGAGGAAG TAAGTTGTAT AGAAACACAG 2400

GACAATCAGA ACTCAGAAGG AGAGGCAAGA AATTGCTGTG AAACATCTAT CAGACAGGAC 2460 TCTGATTCAT CAGTTTCAGA CAAACAGCGG CAAACCATCA TTATTGCCGA CTCCCCGAGT 2520

CCTGCAGTGA GTGTCATCAC TATCAGCAGT GACACTGATG AGGAAGAGAC TCCCCAGAGA 2580

CATTCACTCA GAGAATGTAA AGGTAGTCTA GATTGTGAAG CTTGCCAGAG CACTTTGAAT 2640

ATTGATCGGA TGTGTTCATT AAGTCCTGAT AGTACTCTGC AGTCCAGCCC ATGCCCCTGC 2700

AAGAGACCGA ATAGTATGTC AGATGAAGAG CAAGAAAGTA GTTGTGATAC GGTGGATGGC 2760 TCTCCGACAT CTGACTCTTC CGGGCATGAC AGTCCATTTG CAGAGAGCAC TTTTGTGGAG 2820

GACACTCATG AAAACACAGA ATTGGTATCC TCTGCTGACA CAGAAACCAA GCCAGCTGTC 2880

TGTTCTGTTG TGGTGCCACC AGTGGAACTA GAAAATGGCT TAAATGCCGA TGAGCATATG 2940

GCAAACACAG ATTCTATATG CCAGCCATTA ATAAAAGGAC GATCTTGCCC CTGGAAGATT 3000

AAACCAGCCT TCTGCAGTGG GTACTCGTCA GCAAAAATTG ACATCAGCAT TCCAGCAGCA 3060 GCATTTGAAC TTCAGTCAGG TTCAGCACTT TGGATCTGGG CTACAAGAGT GGAATGGAAA 3120

CTTTGGGCAC AGAAGACAGC AAGCTTATAT TCCTATCGTG TTACCAGTAA TCCATTCACT 3180 CTTTCTCATG GAAGTCCCAA TCACACAGCA GTGCATGCCC ACCTGGCTGG AAATACACAC 3240

CTCGGAGGAC AGCCTACTCT ACTTCCATAC CCATCATCAG CCACCCTCAG TAGTGCTGCA 3300 CCAGTGGCCC ACCTGTTAGC CTCTCCGTGT ACCTCAAGAC CTATGTTACA GCATCCAACT 3360

TATAATATCT CCCATCCCAG TGGCATACTT CACCAAGTCC CAGTGGGCTT AAATCCCCGT 3420

GTGTTACCAT CCCCAACCAT TCATCAGACT CAGTACAAAC CAATCTTCCC ACCACATTCT 3480

TAACATGCAG CAATCACCTG CATATACTGA TTCCACTGAG TCCAACAAAA CTCAGCCAGT 3540

ATCCATATAT GTGAAAAACA GTATATTGGG AAGCTCAATG ATACAAACAT TTGATTAAAA 3600 Note the (TGA)at positions 3592-3594, is the (first) stop codon

ATAAA 3605

(Note the (TGA)at positions 3592-3594, is the (first) stop codon and the remaining sequences probably do not code for amino acids.)

The amino acid sequence, polymer, protein or peptide that corresponds to the above sequence is: Gin Val Leu Val Tyr Pro Pro Tyr Val Tyr Gin Thr Gin Ser Ser Ala 1 5 10 15

Phe Cys Ser Val Lys Lys Leu Lys Val Glu Pro Ser Ser Cys Val Phe 20 25 30

Gin Glu Arg Asn Tyr Pro Arg Thr Tyr Val Asn Gly Arg Asn Phe Gly 35 40 45

Asn Ser His Pro Pro Thr Lys Gly Ser Ala Phe Gin Thr Lys lie Pro 50 55 60

Phe Asn Arg Pro Arg Gly His Asn Phe Ser Leu Gin Thr Ser Ala Val 65 70 75 80 Val Leu Lys Asn Thr Ala Gly Ala Thr Lys Val lie Ala Ala Gin Ala

85 90 95

Gin Gin Ala His Val Gin Ala Pro Gin He Gly Val Trp Arg Asn Arg 100 105 110

Leu His Phe Leu Glu Gly Pro Gin Arg Cys Gly Leu Lys Arg Lys Ser 115 120 125

Glu Glu Leu Asp Asn His Ser Ser Ala Met Gin He Val Asp Glu Leu 130 135 140

Ser He Leu Pro Ala Met Leu Gin Thr Asn Met Glu Asn Pro Val Thr 145 150 155 160 Val Val Thr Ala Thr Thr Gly Ser Lys Gin Asn Cys Thr Thr Gly Glu

165 170 175

Gly Asp Tyr Gin Leu Val Gin His Glu Val Leu Cys Ser Met Lys Asn 180 185 190 Note, Table 3 starts with the aa (tyr) from the sequence at 179. The first "*" in Table 3 would correspond to the position between positions 186 (Leu) and 187 (Cys) .

Thr Tyr Glu Val Leu Asp Phe Val Gly Arg Gly Thr Phe Gly Gin Val 195 200 205

Val Lys Cys Trp Lys Arg Gly Thr Asn Glu lie Val Ala He Lys He 210 215 220 Leu Lys Asn His Pro Ser Tyr Ala Arg Gin Gly Gin He Glu Val Ser

225 230 235 240

He Leu Ala Arg Leu Ser Thr Glu Asn Ala Asp Glu Tyr Asn Phe Val 245 250 255

Arg Ala Tyr Glu Thr Phe Gin His Arg Asn His Thr Cys Leu Val Phe

260 265 270 Glu Met Leu Glu Gin Asn Leu Tyr Asp Phe Leu Lys Gin Asn Lys Phe

275 280 285

Ser Pro Leu Pro Leu Lys Val He Arg Pro He Leu Gin Gin Val Ala 290 295 300

Thr Ala Leu Lys Lys Leu Lys Ser Leu Gly Leu He His Ala Asp Leu 305 310 315 320

Lys Pro Glu Asn He Met Leu Val Asp Pro Val Arg Gin Pro Tyr Arg 325 330 335

Val Lys Val He Asp Phe Gly Ser Ala Ser His Val Ser Lys Thr Val 340 345 350

Trp Ser Thr Tyr Leu Gin Ser Arg Tyr Arg Ala Pro Glu He He Leu 355 360 365

Gly Leu Pro Phe Cys Lys Pro He Asp Met Trp Ser Leu Gly Cys Val 370 375 380

He Ala Glu Leu Phe Leu Gly Trp Pro Leu Tyr Pro Gly Ala Leu Glu 385 390 395 400

Tyr Asp Gin He Arg Tyr He Ser Gin Thr Gin Gly Leu Pro Gly Glu 405 410 415

Gin Leu Leu Asn Val Gly Thr Lys Ser Thr Arg Phe Phe Cys Lys Glu 420 425 430 Thr Asp Met Ser His Ser Gly Trp Arg Leu Lys Thr Leu Glu Glu His 435 440 445

Glu Ala Glu Thr Gly Met Lys Ser Lys Glu Ala Arg Lys Tyr He Phe

450 455 460

Asn Ser Leu Asp Asp Val Ala His Val Asn Thr Val Met Asp Leu Glu

465 470 475 480

Gly Ser Asp Leu Phe Pro Glu Lys Ala Asp Arg Arg Glu Phe Val Ser 485 490 495

Leu Leu Lys Lys Met Leu Leu He Asp Ala Asp Leu Arg He Thr Pro

500 505 510 Ala Glu Thr Leu Asn His Pro Phe Val Asn Met Lys His Leu Leu Asp 515 520 525

Phe Pro His Ser Asn His Val Lys Ser Cys Phe His He Met Asp He 530 535 540

Cys Lys Ser His Leu Asn Ser Cys Asp Thr Asn Asn His Asn Lys Thr 545 550 555 560

Ser Leu Leu Arg Pro Val Ala Ser Ser Ser Thr Ala Thr Leu Thr Ala 565 570 575

Asn Phe Thr Lys He Gly Thr Leu Arg Ser Gin Ala Leu Thr Thr Ser 580 585 590

Ala His Ser Val Val Ala Asp Gly He Pro Leu Gin Ala Gly Thr Arg 595 600 605

Gin Phe Gly Cys Gly Asp Ala Phe Gin Gin Thr Leu He He Cys Pro 610 615 620

Pro Ala He Gin Gly He Pro Ala Thr His Gly Lys Pro Thr Ser Tyr 625 630 635 640

Ser He Arg Val His Asn Thr Val Pro Leu Leu Thr Gin Ala Pro Ala 645 650 655 Val Gin Pro Leu Gin He Arg Pro Arg Ser Ser Phe Leu Arg Arg Pro

•660 665 670

Leu Val Glu His Arg Gin Met Leu Val Pro Ala Trp Gin Gin Val Thr

675 680 685

Pro Leu Ala Pro Ala Thr Thr Thr Leu Thr Ser Glu Ser Val Ala Gly

690 695 700

Ser His Arg Leu Gly Asp Trp Gly Lys Met He Ser Cys Ser Asn His 705 710 715 720

Tyr Asn Ser Val Met Pro Gin Pro Leu Leu Thr Asn Gin He Thr Leu

725 730 735 Ser Ala Pro Gin Pro Val Ser Val Gly He Ala His Val Val Trp Pro

740 745 750

Gin Pro Ala Thr Thr Lys Lys Asn Lys Gin Cys Gin Asn Arg Ser Asn 755 760 765

Ser Leu Gin Asn Thr Asn He Pro Leu Ser Ala Phe He Ser Pro Lys 770 775 780

He He Asn Gly Lys Asp Val Glu Glu Val Ser Cys He Glu Thr Gin 785 790 795 800

Asp Asn Gin Asn Ser Glu Gly Glu Ala Arg Asn Cys Cys Glu Thr Ser 805 810 815

He Arg Gin Asp Ser Asp Ser Ser Val Ser Asp Lys Gin Arg Gin Thr 820 825 830

He He He Ala Asp Ser Pro Ser Pro Ala Val Ser Val He Thr He

835 840 845

Ser Ser Asp Thr Asp Glu Glu Glu Thr Pro Gin Arg His Ser Leu Arg

850 855 860

Glu Cys Lys Gly Ser Leu Asp Cys Glu Ala Cys Gin Ser Thr Leu Asn 865 870 875 880

He Asp Arg Met Cys Ser Leu Ser Pro Asp Ser Thr Leu Gin Ser Ser

885 890 895 Pro Cys Pro Cys Lys Arg Pro Asn Ser Met Ser Asp Glu Glu Gin Glu

900 905 910

Ser Ser Cys Asp Thr Val Asp Gly Ser Pro Thr Ser Asp Ser Ser Gly 915 920 925

His Asp Ser Pro Phe Ala Glu Ser Thr Phe Val Glu Asp Thr His Glu 930 935 940 Asn Thr Glu Leu Val Ser Ser Ala Asp Thr Glu Thr Lys Pro Ala Val 945 950 955 960

Cys Ser Val Val Val Pro Pro Val Glu Leu Glu Asn Gly Leu Asn Ala 965 970 975

Asp Glu His Met Ala Asn Thr Asp Ser He Cys Gin Pro Leu He Lys 980 985 990 Gly Arg Ser Cys Pro Trp Lys He Lys Pro Ala Phe Cys Ser Gly Tyr 995 1000 1005

Ser Ser Ala Lys He Asp He Ser He Pro Ala Ala Ala Phe Glu Leu 1010 1015 1020

Gln Ser Gly Ser Ala Leu Trp He Trp Ala Thr Arg Val Glu Trp Lys 102255 1030 1035 104(

Leu Trp Ala Gin Lys Thr Ala Ser Leu Tyr Ser Tyr Arg Val Thr Ser 1045 1050 1055

Asn Pro Phe Thr Leu Ser His Gly Ser Pro Asn His Thr Ala Val His 1060 1065 1070 Ala His Leu Ala Gly Asn Thr His Leu Gly Gly Gin Pro Thr Leu Leu 1075 1080 1085

Pro Tyr Pro Ser Ser Ala Thr Leu Ser Ser Ala Ala Pro Val Ala His 1090 1095 1100

Leu Leu Ala Ser Pro Cys Thr Ser Arg Pro Met Leu Gin His Pro Thr 1105 1110 1115 1120

Tyr Asn He Ser His Pro Ser Gly He Leu His Gin Val Pro Val Gly 1125 1130 1135

Leu Asn Pro Arg Val Leu Pro Ser Pro Thr He His Gin Thr Gin Tyr 1140 1145 1150

Lys Pro He Phe Pro Pro His Ser His Ala Ala He Thr Cys He Tyr 1155 1160 1165

Phe His Val Gin Gin Asn Ser Ala Ser He His He Cys Glu Lys Gin 1170 1175 1180

Tyr He Gly Lys Leu Asn Asp Thr Asn He @ Leu Lys He 1185 1190 1195

The "@" at position 1195 shows the first stop codon, the following amino acids, Leu Lys He, are probably not part of the functional protein. The sequences in the sequence I.D. listings list all the amino acids, including Leu, Lys, and He.

Table 2 Comparison of 170 kDa kinase (PKR) Activity to PKR mRNA Expression

¹170 kDa kinase activity was estimated from in situ kinase activity blots (Sampson et al., 1993).

+++ is the highest level of activity

++ is a moderate level of activity

+ is a low level of activity - is no measurable activity

²mRNA values were measured by determining radioactive counts emitted from Northern blots by an AMBIS beta-counter. The numbers provided represent the average of two blots given in counts. Numbers in parenthesis represent the ratio of PKR levels in the resistant cell line compared to the levels in the respective sensitive cell line.

Table 3

Comparison of Amino Acid Sequence of a selected segment of PKR to selected segments of Yakl,Cdk ,PKA and PKC. Amino acids below in the same position as PKR amino acids can be substituted for the same position PKR amino acids from Yakl, Cdk4, PKA or PKC (or added to the PKR sequence If marked with an " * . " )

SEQUENCE LISTING

(1) GENERAL INFORMATION:

(i) APPLICANT: Upjohn Company, The

(ii) TITLE OF INVENTION: A NOVEL GENE FOR A PROTEIN KINASE ASSOCIATED WITH DRUG RESISTANCE

(iii) NUMBER OF SEQUENCES: 13

(iv) CORRESPONDENCE ADDRESS:

(A) ADDRESSEE: The Upjohn Company, Intellect. Prop. Law (1920-32-1)

(B) STREET: 301 Henrietta Street

(C) CITY: Kalamazoo

(D) STATE: Michigan

(E) COUNTRY: U.S.A. (F) ZIP: 49001

(V) COMPUTER READABLE FORM:

(A) MEDIUM TYPE: Floppy disk

(B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS

(D) SOFTWARE: Patentln Release #1.0, Version #1.25

(vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: (B) FILING DATE:

(C) CLASSIFICATION:

(viii) ATTORNEY/AGENT INFORMATION:

(A) NAME: Wootton, Thomas A. (B) REGISTRATION NUMBER: 35,004

(C) REFERENCE/DOCKET NUMBER: 4879.P CP

(ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: (616) 385-7914 (B) TELEFAX: (616) 385-6897

(C) TELEX: 224401

(2) INFORMATION FOR SEQ ID NO:l:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 3594 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: CDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(Xi) SEQUENCE DESCRIPTION: SEQ ID NO:l:

CAAGTCTTGG TCTACCCACC ATATGTTTAT CAAACTCAGT CAAGTGCCTT TTGTAGTGTG 60

AAGAAACTCA AAGTAGAGCC AAGCAGTTGT GTATTCCAGG AAAGAAACTA TCCACGGACC 120

TATGTGAATG GTAGAAACTT TGGAAATTCT CATCCTCCCA CTAAGGGTAG TGCTTTTCAG 180 ACAAAGATAC CATTTAATAG ACCTCGAGGA CACAACTTTT CATTGCAGAC AAGTGCTGTT 240

GTTTTGAAAA ACACTGCAGG TGCTACAAAG GTCATAGCAG CTCAGGCACA GCAAGCTCAC 300 GTGCAGGCCC CTCAGATTGG GGTGTGGCGA AACAGATTGC ATTTCCTAGA AGGCCCCCAG 360

CGATGTGGAT TGAAGCGCAA GAGTGAGGAG TTGGATAATC ATAGCAGCGC AATGCAGATT 420

GTCGATGAAT TGTCCATACT TCCTGCAATG TTGCAAACCA ACATGGAAAA TCCAGTGACA 480

GTTGTGACAG CTACCACAGG ATCAAAACAG AATTGTACCA CTGGAGAAGG TGACTATCAG 540

TTAGTACAGC ATGAAGTCTT ATGCTCCATG AAAAATACTT ACGAAGTCCT TGATTTTGTT 600 GGTCGAGGCA CGTTTGGCCA GGTAGTTAAA TGCTGGAAAA GAGGGACAAA TGAAATTGTA 660

GCAATCAAAA TTTTGAAGAA TCATCCTTCT TATGCCCGTC AAGGTCAAAT AGAAGTGAGC 720

ATATTAGCAA GGCTCAGTAC TGAAAATGCT GATGAATATA ACTTTGTACG AGCTTATGAA 780

ACGTTTCAGC ACCGTAACCA TACTTGTTTA GTCTTTGAGA TGCTGGAACA AAACTTGTAT 840

GACTTTCTGA AACAAAATAA ATTTAGTCCC CTGCCTCTAA AAGTGATTCG GCCCATTCTT 900 CAACAAGTGG CCACTGCACT GAAAAAATTG AAAAGTCTTG GTTTAATTCA TGCTGATCTA 960

AAGCCTGAGA ATATTATGTT GGTGGATCCT GTTCGGCAGC CTTACAGGGT TAAAGTAATA 1020

GACTTTGGGT CGGCCAGTCA TGTATCAAAG ACTGTTTGGT CAACATATCT ACAATCTCGG 1080

TACAGAGCTC CAGAGATTAT ATTGGGGTTG CCATTTTGTA AGCCAATAGA CATGTGGTCA 1140

TTGGGATGTG TGATTGCAGA ATTATTTCTT GGATGGCCGC TATACCCAGG AGCCTTGGAG 1200 TATGATCAGA TTCGATACAT TTCTCAGACT CAAGGTTTGC CAGGAGAACA GTTGTTAAAT 1260

GTGGGTACTA AATCCACAAG ATTTTTTTGC AAAGAAACAG ATATGTCTCA TTCTGGTTGG 1320

AGATTAAAGA CATTGGAAGA GCATGAGGCA GAGACAGGAA TGAAGTCTAA AGAAGCCAGA 1380

AAATACATTT TCAACAGTCT GGATGATGTA GCGCATGTGA ACACAGTGAT GGATTTGGAA 1440

GGAAGTGATC TTTTTCCTGA GAAAGCTGAT AGAAGAGAAT TTGTTAGTCT GTTGAAGAAA 1500 ATGTTGCTGA TTGATGCAGA TTTAAGAATT ACTCCAGCTG AGACCCTGAA CCATCCTTTT 1560

GTTAATATGA AACATCTTCT AGATTTCCCT CATAGCAACC ATGTAAAGTC CTGTTTTCAT 1620

ATTATGGATA TTTGTAAGTC CCACCTAAAT TCATGTGACA CAAATAATCA CAACAAAACT 1680

TCACTTTTAA GACCAGTTGC TTCAAGCAGT ACTGCTACAC TGACTGCAAA TTTTACTAAA 1740

ATCGGAACAT TAAGAAGTCA GGCATTGACC ACATCTGCTC ATTCAGTTGT GGCGGATGGA 1800 ATACCTCTGC AGGCAGGAAC TCGTCAGTTT GGTTGTGGTG ATGCTTTTCA GCAGACATTG 1860

ATTATCTGTC CCCCAGCTAT TCAAGGTATT CCTGCAACAC ATGGTAAACC CACCAGTTAT 1920

TCAATAAGGG TACATAATAC AGTTCCACTT CTAACTCAGG CCCCAGCTGT GCAGCCACTA 1980

CAGATCCGAC CAAGGAGTTC TTTTCTCAGA CGTCCTCTGG TAGAACACAG ACAGATGCTG 2040

GTGCCTGCCT GGCAACAGGT GACACCCCTG GCTCCTGCTA CTACTACACT AACTTCTGAG 2100 AGTGTGGCTG GTTCACACAG GCTTGGAGAC TGGGGGAAGA TGATTTCATG CAGCAATCAT 2160

TATAACTCAG TGATGCCGCA GCCTCTTCTG ACCAATCAGA TAACTTTATC TGCCCCTCAG 2220 CCAGTTAGTG TGGGGATTGC ACATGTTGTC TGGCCTCAGC CTGCCACTAC CAAGAAAAAT 2280

AAACAGTGCC AGAACAGGAG TAATTCATTA CAGAATACCA ATATCCCACT ATCAGCATTT 2340 ATTTCTCCAA AGATAATTAA TGGGAAAGAT GTCGAGGAAG TAAGTTGTAT AGAAACACAG 2400

GACAATCAGA ACTCAGAAGG AGAGGCAAGA AATTGCTGTG AAACATCTAT CAGACAGGAC 2460

TCTGATTCAT CAGTTTCAGA CAAACAGCGG CAAACCATCA TTATTGCCGA CTCCCCGAGT 2520

CCTGCAGTGA GTGTCATCAC TATCAGCAGT GACACTGATG AGGAAGAGAC TCCCCAGAGA 2580

CATTCACTCA GAGAATGTAA AGGTAGTCTA GATTGTGAAG CTTGCCAGAG CACTTTGAAT 2640 ATTGATCGGA TGTGTTCATT AAGTCCTGAT AGTACTCTGC AGTCCAGCCC ATGCCCCTGC 2700

AAGAGACCGA ATAGTATGTC AGATGAAGAG CAAGAAAGTA GTTGTGATAC GGTGGATGGC 2760

TCTCCGACAT CTGACTCTTC CGGGCATGAC AGTCCATTTG CAGAGAGCAC TTTTGTGGAG 2820

GACACTCATG AAAACACAGA ATTGGTATCC TCTGCTGACA CAGAAACCAA GCCAGCTGTC 2880

TGTTCTGTTG TGGTGCCACC AGTGGAACTA GAAAATGGCT TAAATGCCGA TGAGCATATG 2940 GCAAACACAG ATTCTATATG CCAGCCATTA ATAAAAGGAC GATCTTGCCC CTGGAAGATT 3000

AAACCAGCCT TCTGCAGTGG GTACTCGTCA GCAAAAATTG ACATCAGCAT TCCAGCAGCA 3060

GCATTTGAAC TTCAGTCAGG TTCAGCACTT TGGATCTGGG CTACAAGAGT GGAATGGAAA 3120

CTTTGGGCAC AGAAGACAGC AAGCTTATAT TCCTATCGTG TTACCAGTAA TCCATTCACT 3180

CTTTCTCATG GAAGTCCCAA TCACACAGCA GTGCATGCCC ACCTGGCTGG AAATACACAC 3240 CTCGGAGGAC AGCCTACTCT ACTTCCATAC CCATCATCAG CCACCCTCAG TAGTGCTGCA 3300

CCAGTGGCCC ACCTGTTAGC CTCTCCGTGT ACCTCAAGAC CTATGTTACA GCATCCAACT 3360

TATAATATCT CCCATCCCAG TGGCATACTT CACCAAGTCC CAGTGGGCTT AAATCCCCGT 3420

GTGTTACCAT CCCCAACCAT TCATCAGACT CAGTACAAAC CAATCTTCCC ACCACATTCT 3480

TAACATGCAG CAATCACCTG CATATACTGA TTCCACTGAG TCCAACAAAA CTCAGCCAGT 3540 ATCCATATAT GTGAAAAACA GTATATTGGG AAGCTCAATG ATACAAACAT TTGA 3594

(2) INFORMATION FOR SEQ ID NO:2:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 3060 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: CDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:2 : TATCAGTTAG TACAGCATGA AGTCTTATGC TCCATGAAAA ATACTTACGA AGTCCTTGAT 60

TTTGTTGGTC GAGGCACGTT TGGCCAGGTA GTTAAATGCT GGAAAAGAGG GACAAATGAA 120 ATTGTAGCAA TCAAAATTTT GAAGAATCAT CCTTCTTATG CCCGTCAAGG TCAAATAGAA 180

GTGAGCATAT TAGCAAGGCT CAGTACTGAA AATGCTGATG AATATAACTT TGTACGAGCT 240 TATGAAACGT TTCAGCACCG TAACCATACT TGTTTAGTCT TTGAGATGCT GGAACAAAAC 300

TTGTATGACT TTCTGAAACA AAATAAATTT AGTCCCCTGC CTCTAAAAGT GATTCGGCCC 360

ATTCTTCAAC AAGTGGCCAC TGCACTGAAA AAATTGAAAA GTCTTGGTTT AATTCATGCT 420

GATCTAAAGC CTGAGAATAT TATGTTGGTG GATCCTGTTC GGCAGCCTTA CAGGGTTAAA 480

GTAATAGACT TTGGGTCGGC CAGTCATGTA TCAAAGACTG TTTGGTCAAC ATATCTACAA 540 TCTCGGTACA GAGCTCCAGA GATTATATTG GGGTTGCCAT TTTGTAAGCC AATAGACATG 600

TGGTCATTGG GATGTGTGAT TGCAGAATTA TTTCTTGGAT GGCCGCTATA CCCAGGAGCC 660

TTGGAGTATG ATCAGATTCG ATACATTTCT CAGACTCAAG GTTTGCCAGG AGAACAGTTG 720

TTAAATGTGG GTACTAAATC CACAAGATTT TTTTGCAAAG AAACAGATAT GTCTCATTCT 780

GGTTGGAGAT TAAAGACATT GGAAGAGCAT GAGGCAGAGA CAGGAATGAA GTCTAAAGAA 840 GCCAGAAAAT ACATTTTCAA CAGTCTGGAT GATGTAGCGC ATGTGAACAC AGTGATGGAT 900

TTGGAAGGAA GTGATCTTTT TCCTGAGAAA GCTGATAGAA GAGAATTTGT TAGTCTGTTG 960

AAGAAAATGT TGCTGATTGA TGCAGATTTA AGAATTACTC CAGCTGAGAC CCTGAACCAT 1020

CCTTTTGTTA ATATGAAACA TCTTCTAGAT TTCCCTCATA GCAACCATGT AAAGTCCTGT 1080

TTTCATATTA TGGATATTTG TAAGTCCCAC CTAAATTCAT GTGACACAAA TAATCACAAC 1140 AAAACTTCAC TTTTAAGACC AGTTGCTTCA AGCAGTACTG CTACACTGAC TGCAAATTTT 1200

ACTAAAATCG GAACATTAAG AAGTCAGGCA TTGACCACAT CTGCTCATTC AGTTGTGGCG 1260

GATGGAATAC CTCTGCAGGC AGGAACTCGT CAGTTTGGTT GTGGTGATGC TTTTCAGCAG 1320

ACATTGATTA TCTGTCCCCC AGCTATTCAA GGTATTCCTG CAACACATGG TAAACCCACC 1380

AGTTATTCAA TAAGGGTACA TAATACAGTT CCACTTCTAA CTCAGGCCCC AGCTGTGCAG 1440 CCACTACAGA TCCGACCAAG GAGTTCTTTT CTCAGACGTC CTCTGGTAGA ACACAGACAG 1500

ATGCTGGTGC CTGCCTGGCA ACAGGTGACA CCCCTGGCTC CTGCTACTAC TACACTAACT 1560

TCTGAGAGTG TGGCTGGTTC ACACAGGCTT GGAGACTGGG GGAAGATGAT TTCATGCAGC 1620

AATCATTATA ACTCAGTGAT GCCGCAGCCT CTTCTGACCA ATCAGATAAC TTTATCTGCC 1680

CCTCAGCCAG TTAGTGTGGG GATTGCACAT GTTGTCTGGC CTCAGCCTGC CACTACCAAG 1740 AAAAATAAAC AGTGCCAGAA CAGGAGTAAT TCATTACAGA ATACCAATAT CCCACTATCA 1800

GCATTTATTT CTCCAAAGAT AATTAATGGG AAAGATGTCG AGGAAGTAAG TTGTATAGAA 1860

ACACAGGACA ATCAGAACTC AGAAGGAGAG GCAAGAAATT GCTGTGAAAC ATCTATCAGA 1920

CAGGACTCTG ATTCATCAGT TTCAGACAAA CAGCGGCAAA CCATCATTAT TGCCGACTCC 1980

CCGAGTCCTG CAGTGAGTGT CATCACTATC AGCAGTGACA CTGATGAGGA AGAGACTCCC 2040 CAGAGACATT CACTCAGAGA ATGTAAAGGT AGTCTAGATT GTGAAGCTTG CCAGAGCACT 2100

TTGAATATTG ATCGGATGTG TTCATTAAGT CCTGATAGTA CTCTGCAGTC CAGCCCATGC 2160 CCCTGCAAGA GACCGAATAG TATGTCAGAT GAAGAGCAAG AAAGTAGTTG TGATACGGTG 2220

GATGGCTCTC CGACATCTGA CTCTTCCGGG CATGACAGTC CATTTGCAGA GAGCACTTTT 2280 GTGGAGGACA CTCATGAAAA CACAGAATTG GTATCCTCTG CTGACACAGA AACCAAGCCA 2340

GCTGTCTGTT CTGTTGTGGT GCCACCAGTG GAACTAGAAA ATGGCTTAAA TGCCGATGAG 2400

CATATGGCAA ACACAGATTC TATATGCCAG CCATTAATAA AAGGACGATC TTGCCCCTGG 2460

AAGATTAAAC CAGCCTTCTG CAGTGGGTAC TCGTCAGCAA AAATTGACAT CAGCATTCCA 2520

GCAGCAGCAT TTGAACTTCA GTCAGGTTCA GCACTTTGGA TCTGGGCTAC AAGAGTGGAA 2580 TGGAAACTTT GGGCACAGAA GACAGCAAGC TTATATTCCT ATCGTGTTAC CAGTAATCCA 2640

TTCACTCTTT CTCATGGAAG TCCCAATCAC ACAGCAGTGC ATGCCCACCT GGCTGGAAAT 2700

ACACACCTCG GAGGACAGCC TACTCTACTT CCATACCCAT CATCAGCCAC CCTCAGTAGT 2760

GCTGCACCAG TGGCCCACCT GTTAGCCTCT CCGTGTACCT CAAGACCTAT GTTACAGCAT 2820

CCAACTTATA ATATCTCCCA TCCCAGTGGC ATACTTCACC AAGTCCCAGT GGGCTTAAAT 2880 CCCCGTGTGT TACCATCCCC AACCATTCAT CAGACTCAGT ACAAACCAAT CTTCCCACCA 2940

CATTCTTAAC ATGCAGCAAT CACCTGCATA TACTGATTCC ACTGAGTCCA ACAAAACTCA 3000

GCCAGTATCC ATATATGTGA AAAACAGTAT ATTGGGAAGC TCAATGATAC AAACATTTGA 3060

(2) INFORMATION FOR SEQ ID NO: 3 :

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 729 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3: TATCAGTTAG TACAGCATGA AGTCTTATGC TCCATGAAAA ATACTTACGA AGTCCTTGAT 60

TTTGTTGGTC GAGGCACGTT TGGCCAGGTA GTTAAATGCT GGAAAAGAGG GACAAATGAA 120

ATTGTAGCAA TCAAAATTTT GAAGAATCAT CCTTCTTATG CCCGTCAAGG TCAAATAGAA 180

GTGAGCATAT TAGCAAGGCT CAGTACTGAA AATGCTGATG AATATAACTT TGTACGAGCT 240

TATGAAACGT TTCAGCACCG TAACCATACT TGTTTAGTCT TTGAGATGCT GGAACAAAAC 300 TTGTATGACT TTCTGAAACA AAATAAATTT AGTCCCCTGC CTCTAAAAGT GATTCGGCCC 360

ATTCTTCAAC AAGTGGCCAC TGCACTGAAA AAATTGAAAA GTCTTGGTTT AATTCATGCT 420

GATCTAAAGC CTGAGAATAT TATGTTGGTG GATCCTGTTC GGCAGCCTTA CAGGGTTAAA 480

GTAATAGACT TTGGGTCGGC CAGTCATGTA TCAAAGACTG TTTGGTCAAC ATATCTACAA 540 TCTCGGTACA GAGCTCCAGA GATTATATTG GGGTTGCCAT TTTGTAAGCC AATAGACATG 600

TGGTCATTGG GATGTGTGAT TGCAGAATTA TTTCTTGGAT GGCCGCTATA CCCAGGAGCC 660 TTGGAGTATG ATCAGATTCG ATACATTTCT CAGACTCAAG GTTTGCCAGG AGAACAGTTG 720 TTAAATGTG 729

(2) INFORMATION FOR SEQ ID NO: 4 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1194 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: single (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: N-terminal

(Xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:

Gin Val Leu Val Tyr Pro Pro Tyr Val Tyr Gin Thr Gin Ser Ser Ala 1 5 10 15

Phe Cys Ser Val Lys Lys Leu Lys Val Glu Pro Ser Ser Cys Val Phe

20 25 30 Gin Glu Arg Asn Tyr Pro Arg Thr Tyr Val Asn Gly Arg Asn Phe Gly 35 40 45

Asn Ser His Pro Pro Thr Lys Gly Ser Ala Phe Gin Thr Lys He Pro 50 55 60

Phe Asn Arg Pro Arg Gly His Asn Phe Ser Leu Gin Thr Ser Ala Val 65 70 75 80

Val Leu Lys Asn Thr Ala Gly Ala Thr Lys Val He Ala Ala Gin Ala 85 90 95

Gin Gin Ala His Val Gin Ala Pro Gin He Gly Val Trp Arg Asn Arg 100 105 110

Leu His Phe Leu Glu Gly Pro Gin Arg Cys Gly Leu Lys Arg Lys Ser 115 120 125

Glu Glu Leu Asp Asn His Ser Ser Ala Met Gin He Val Asp Glu Leu 130 135 140

Ser He Leu Pro Ala Met Leu Gin Thr Asn Met Glu Asn Pro Val Thr 145 150 155 160

Val Val Thr Ala Thr Thr Gly Ser Lys Gin Asn Cys Thr Thr Gly Glu 165 170 175

Gly Asp Tyr Gin Leu Val Gin His Glu Val Leu Cys Ser Met Lys Asn 180 185 190 Thr Tyr Glu Val Leu Asp Phe Val Gly Arg Gly Thr Phe Gly Gin Val 195 200 205 Val Lys Cys Trp Lys Arg Gly Thr Asn Glu He Val Ala He Lys He

210 215 220

Leu Lys Asn His Pro Ser Tyr Ala Arg Gin Gly Gin He Glu Val Ser 225 230 235 240

He Leu Ala Arg Leu Ser Thr Glu Asn Ala Asp Glu Tyr Asn Phe Val

245 250 255 Arg Ala Tyr Glu Thr Phe Gin His Arg Asn His Thr Cys Leu Val Phe

260 265 270

Glu Met Leu Glu Gin Asn Leu Tyr Asp Phe Leu Lys Gin Asn Lys Phe 275 280 285

Ser Pro Leu Pro Leu Lys Val He Arg Pro He Leu Gin Gin Val Ala 290 295 300

Thr Ala Leu Lys Lys Leu Lys Ser Leu Gly Leu He His Ala Asp Leu 305 310 315 320

Lys Pro Glu Asn He Met Leu Val Asp Pro Val Arg Gin Pro Tyr Arg 325 330 335

Val Lys Val He Asp Phe Gly Ser Ala Ser His Val Ser Lys Thr Val 340 345 350

Trp Ser Thr Tyr Leu Gin Ser Arg Tyr Arg Ala Pro Glu He He Leu

355 360 365

Gly Leu Pro Phe Cys Lys Pro He Asp Met Trp Ser Leu Gly Cys Val

370 375 380

He Ala Glu Leu Phe Leu Gly Trp Pro Leu Tyr Pro Gly Ala Leu Glu 385 390 395 400

Tyr Asp Gin He Arg Tyr He Ser Gin Thr Gin Gly Leu Pro Gly Glu

405 410 415 Gin Leu Leu Asn Val Gly Thr Lys Ser Thr Arg Phe Phe Cys Lys Glu

420 425 430

Thr Asp Met Ser His Ser Gly Trp Arg Leu Lys Thr Leu Glu Glu His 435 440 445

Glu Ala Glu Thr Gly Met Lys Ser Lys Glu Ala Arg Lys Tyr He Phe 450 455 460

Asn Ser Leu Asp Asp Val Ala His Val Asn Thr Val Met Asp Leu Glu 465 470 475 480

Gly Ser Asp Leu Phe Pro Glu Lys Ala Asp Arg Arg Glu Phe Val Ser

485 490 495 Leu Leu Lys Lys Met Leu Leu He Asp Ala Asp Leu Arg He Thr Pro

500 505 510

Ala Glu Thr Leu Asn His Pro Phe Val Asn Met Lys His Leu Leu Asp 515 520 525

Phe Pro His Ser Asn His Val Lys Ser Cys Phe His He Met Asp He 530 535 540

Cys Lys Ser His Leu Asn Ser Cys Asp Thr Asn Asn His Asn Lys Thr 545 550 555 560

Ser Leu Leu Arg Pro Val Ala Ser Ser Ser Thr Ala Thr Leu Thr Ala 565 570 575

Asn Phe Thr Lys He Gly Thr Leu Arg Ser Gin Ala Leu Thr Thr Ser 580 585 590

Ala His Ser Val Val Ala Asp Gly He Pro Leu Gin Ala Gly Thr Arg 595 600 605

Gin Phe Gly Cys Gly Asp Ala Phe Gin Gin Thr Leu He He Cys Pro 610 615 620

Pro Ala He Gin Gly He Pro Ala Thr His Gly Lys Pro Thr Ser Tyr 625 630 635 640 Ser He Arg Val His Asn Thr Val Pro Leu Leu Thr Gin Ala Pro Ala

645 650 655

Val Gin Pro Leu Gin He Arg Pro Arg Ser Ser Phe Leu Arg Arg Pro 660 665 670

Leu Val Glu His Arg Gin Met Leu Val Pro Ala Trp Gin Gin Val Thr 675 680 685

Pro Leu Ala Pro Ala Thr Thr Thr Leu Thr Ser Glu Ser Val Ala Gly 690 695 700

Ser His Arg Leu Gly Asp Trp Gly Lys Met He Ser Cys Ser Asn His 705 710 715 720 Tyr Asn Ser Val Met Pro Gin Pro Leu Leu Thr Asn Gin He Thr Leu

725 730 735

Ser Ala Pro Gin Pro Val Ser Val Gly He Ala His Val Val Trp Pro 740 745 750

Gin Pro Ala Thr Thr Lys Lys Asn Lys Gin Cys Gin Asn Arg Ser Asn 755 760 765

Ser Leu Gin Asn Thr Asn He Pro Leu Ser Ala Phe He Ser Pro Lys 770 775 780

He He Asn Gly Lys Asp Val Glu Glu Val Ser Cys He Glu Thr Gin 785 790 795 800

Asp Asn Gin Asn Ser Glu Gly Glu Ala Arg Asn Cys Cys Glu Thr Ser 805 810 815

He Arg Gin Asp Ser Asp Ser Ser Val Ser Asp Lys Gin Arg Gin Thr

820 825 830

He He He Ala Asp Ser Pro Ser Pro Ala Val Ser Val He Thr He

835 840 845

Ser Ser Asp Thr Asp Glu Glu Glu Thr Pro Gin Arg His Ser Leu Arg 850 855 860

Glu Cys Lys Gly Ser Leu Asp Cys Glu Ala Cys Gin Ser Thr Leu Asn

865 870 875 880 He Asp Arg Met Cys Ser Leu Ser Pro Asp Ser Thr Leu Gin Ser Ser

885 890 895

Pro Cys Pro Cys Lys Arg Pro Asn Ser Met Ser Asp Glu Glu Gin Glu

900 905 910

Ser Ser Cys Asp Thr Val Asp Gly Ser Pro Thr Ser Asp Ser Ser Gly 915 920 925 His Asp Ser Pro Phe Ala Glu Ser Thr Phe Val Glu Asp Thr His Glu 930 935 940

Asn Thr Glu Leu Val Ser Ser Ala Asp Thr Glu Thr Lys Pro Ala Val 945 950 955 960

Cys Ser Val Val Val Pro Pro Val Glu Leu Glu Asn Gly Leu Asn Ala 965 970 975 Asp Glu His Met Ala Asn Thr Asp Ser He Cys Gin Pro Leu He Lys

980 985 990

Gly Arg Ser Cys Pro Trp Lys He Lys Pro Ala Phe Cys Ser Gly Tyr 995 1000 1005

Ser Ser Ala Lys He Asp He Ser He Pro Ala Ala Ala Phe Glu Leu 1010 1015 1020

Gin Ser Gly Ser Ala Leu Trp He Trp Ala Thr Arg Val Glu Trp Lys 1025 1030 1035 1040

Leu Trp Ala Gin Lys Thr Ala Ser Leu Tyr Ser Tyr Arg Val Thr Ser 1045 1050 1055

Asn Pro Phe Thr Leu Ser His Gly Ser Pro Asn His Thr Ala Val His 1060 1065 1070

Ala His Leu Ala Gly Asn Thr His Leu Gly Gly Gin Pro Thr Leu Leu 1075 1080 1085

Pro Tyr Pro Ser Ser Ala Thr Leu Ser Ser Ala Ala Pro Val Ala His 1090 1095 1100

Leu Leu Ala Ser Pro Cys Thr Ser Arg Pro Met Leu Gin His Pro Thr 1105 1110 1115 1120

Tyr Asn He Ser His Pro Ser Gly He Leu His Gin Val Pro Val Gly 1125 1130 1135 Leu Asn Pro Arg Val Leu Pro Ser Pro Thr He His Gin Thr Gin Tyr

1140 1145 1150

Lys Pro He Phe Pro Pro His Ser His Ala Ala He Thr Cys He Tyr 1155 1160 1165

Phe His Val Gin Gin Asn Ser Ala Ser He His He Cys Glu Lys Gin

1170 1175 1180

Tyr He Gly Lys Leu Asn Asp Thr Asn He 1185 1190

(2) INFORMATION FOR SEQ ID NO: 5:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1016 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(v) FRAGMENT TYPE: N-terminal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5 :

Tyr Gin Leu Val Gin His Glu Val Leu Cys Ser Met Lys Asn Thr Tyr 1 5 10 15

Glu Val Leu Asp Phe Val Gly Arg Gly Thr Phe Gly Gin Val Val Lys 20 25 30 Cys Trp Lys Arg Gly Thr Asn Glu He Val Ala He Lys He Leu Lys 35 40 45

Asn His Pro Ser Tyr Ala Arg Gin Gly Gin He Glu Val Ser He Leu 50 55 60

Ala Arg Leu Ser Thr Glu Asn Ala Asp Glu Tyr Asn Phe Val Arg Ala 65 70 75 80

Tyr Glu Thr Phe Gin His Arg Asn His Thr Cys Leu Val Phe Glu Met 85 90 95

Leu Glu Gin Asn Leu Tyr Asp Phe Leu Lys Gin Asn Lys Phe Ser Pro 100 105 110 Leu Pro Leu Lys Val He Arg Pro He Leu Gin Gin Val Ala Thr Ala 115 120 125

Leu Lys Lys Leu Lys Ser Leu Gly Leu He His Ala Asp Leu Lys Pro 130 135 140

Glu Asn He Met Leu Val Asp Pro Val Arg Gin Pro Tyr Arg Val Lys 145 150 155 160

Val He Asp Phe Gly Ser Ala Ser His Val Ser Lys Thr Val Trp Ser 165 170 175

Thr Tyr Leu Gin Ser Arg Tyr Arg Ala Pro Glu He He Leu Gly Leu

180 185 190 Pro Phe Cys Lys Pro He Asp Met Trp Ser Leu Gly Cys Val He Ala

195 200 205

Glu Leu Phe Leu Gly Trp Pro Leu Tyr Pro Gly Ala Leu Glu Tyr Asp 210 215 220

Gin He Arg Tyr He Ser Gin Thr Gin Gly Leu Pro Gly Glu Gin Leu 225 230 235 240

Leu Asn Val Gly Thr Lys Ser Thr Arg Phe Phe Cys Lys Glu Thr Asp 245 250 255

Met Ser His Ser Gly Trp Arg Leu Lys Thr Leu Glu Glu His Glu Ala

260 265 270 Glu Thr Gly Met Lys Ser Lys Glu Ala Arg Lys Tyr He Phe Asn Ser

275 280 285

Leu Asp Asp Val Ala His Val Asn Thr Val Met Asp Leu Glu Gly Ser 290 295 300

Asp Leu Phe Pro Glu Lys Ala Asp Arg Arg Glu Phe Val Ser Leu Leu 305 310 315 320

Lys Lys Met Leu Leu He Asp Ala Asp Leu Arg He Thr Pro Ala Glu 325 330 335

Thr Leu Asn His Pro Phe Val Asn Met Lys His Leu Leu Asp Phe Pro 340 345 350

His Ser Asn His Val Lys Ser Cys Phe His He Met Asp He Cys Lys 355 360 365

Ser His Leu Asn Ser Cys Asp Thr Asn Asn His Asn Lys Thr Ser Leu 370 375 380

Leu Arg Pro Val Ala Ser Ser Ser Thr Ala Thr Leu Thr Ala Asn Phe 385 390 395 400

Thr Lys He Gly Thr Leu Arg Ser Gin Ala Leu Thr Thr Ser Ala His 405 410 415 Ser Val Val Ala Asp Gly He Pro Leu Gin Ala Gly Thr Arg Gin Phe

420 425 430

Gly Cys Gly Asp Ala Phe Gin Gin Thr Leu He He Cys Pro Pro Ala 435 440 445

He Gin Gly He Pro Ala Thr His Gly Lys Pro Thr Ser Tyr Ser He 450 455 460

Arg Val His Asn Thr Val Pro Leu Leu Thr Gin Ala Pro Ala Val Gin 465 470 475 480

Pro Leu Gin He Arg Pro Arg Ser Ser Phe Leu Arg Arg Pro Leu Val

485 490 495 Glu His Arg Gin Met Leu Val Pro Ala Trp Gin Gin Val Thr Pro Leu

500 505 510

Ala Pro Ala Thr Thr Thr Leu Thr Ser Glu Ser Val Ala Gly Ser His 515 520 525

Arg Leu Gly Asp Trp Gly Lys Met He Ser Cys Ser Asn His Tyr Asn 530 535 540

Ser Val Met Pro Gin Pro Leu Leu Thr Asn Gin He Thr Leu Ser Ala 545 550 555 560

Pro Gin Pro Val Ser Val Gly He Ala His Val Val Trp Pro Gin Pro

565 570 575 Ala Thr Thr Lys Lys Asn Lys Gin Cys Gin Asn Arg Ser Asn Ser Leu

580 585 590

Gin Asn Thr Asn He Pro Leu Ser Ala Phe He Ser Pro Lys He He 595 600 605

Asn Gly Lys Asp Val Glu Glu Val Ser Cys He Glu Thr Gin Asp Asn 610 615 620

Gin Asn Ser Glu Gly Glu Ala Arg Asn Cys Cys Glu Thr Ser He Arg 625 630 635 640

Gin Asp Ser Asp Ser Ser Val Ser Asp Lys Gin Arg Gin Thr He He 645 650 655

He Ala Asp Ser Pro Ser Pro Ala Val Ser Val He Thr He Ser Ser 660 665 670

Asp Thr Asp Glu Glu Glu Thr Pro Gin Arg His Ser Leu Arg Glu Cys 675 680 685

Lys Gly Ser Leu Asp Cys Glu Ala Cys Gin Ser Thr Leu Asn He Asp 690 695 700 Arg Met Cys Ser Leu Ser Pro Asp Ser Thr Leu Gin Ser Ser Pro Cys

705 710 715 720

Pro Cys Lys Arg Pro Asn Ser Met Ser Asp Glu Glu Gin Glu Ser Ser 725 730 735

Cys Asp Thr Val Asp Gly Ser Pro Thr Ser Asp Ser Ser Gly His Asp

740 745 750 Ser Pro Phe Ala Glu Ser Thr Phe Val Glu Asp Thr His Glu Asn Thr

755 760 765

Glu Leu Val Ser Ser Ala Asp Thr Glu Thr Lys Pro Ala Val Cys Ser 770 775 780

Val Val Val Pro Pro Val Glu Leu Glu Asn Gly Leu Asn Ala Asp Glu 785 790 795 800

His Met Ala Asn Thr Asp Ser He Cys Gin Pro Leu He Lys Gly Arg 805 810 815

Ser Cys Pro Trp Lys He Lys Pro Ala Phe Cys Ser Gly Tyr Ser Ser 820 825 830 Ala Lys He Asp He Ser He Pro Ala Ala Ala Phe Glu Leu Gin Ser 835 840 845

Gly Ser Ala Leu Trp He Trp Ala Thr Arg Val Glu Trp Lys Leu Trp 850 855 860

Ala Gin Lys Thr Ala Ser Leu Tyr Ser Tyr Arg Val Thr Ser Asn Pro 865 870 875 880

Phe Thr Leu Ser His Gly Ser Pro Asn His Thr Ala Val His Ala His 885 890 895

Leu Ala Gly Asn Thr His Leu Gly Gly Gin Pro Thr Leu Leu Pro Tyr 900 905 910

Pro Ser Ser Ala Thr Leu Ser Ser Ala Ala Pro Val Ala His Leu Leu 915 920 925

Ala Ser Pro Cys Thr Ser Arg Pro Met Leu Gin His Pro Thr Tyr Asn 930 935 940

He Ser His Pro Ser Gly He Leu His Gin Val Pro Val Gly Leu Asn 945 950 955 960

Pro Arg Val Leu Pro Ser Pro Thr He His Gin Thr Gin Tyr Lys Pro 965 970 975

He Phe Pro Pro His Ser His Ala Ala He Thr Cys He Tyr Phe His 980 985 990 Val Gin Gin Asn Ser Ala Ser He His He Cys Glu Lys Gin Tyr He 995 1000 1005

Gly Lys Leu Asn Asp Thr Asn He 1010 1015

(2) INFORMATION FOR SEQ ID NO:6:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 221 amino acids (B) TYPE: amino acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: N-terminal

(Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6:

Tyr Gin Leu Val Gin His Glu Val Leu Cys Ser Met Lys Asn Thr Tyr 1 5 10 15

Glu Val Leu Asp Phe Val Gly Arg Gly Thr Phe Gly Gin Val Val Lys 20 25 30

Cys Trp Lys Arg Gly Thr Asn Glu He Val Ala He Lys He Leu Lys 35 40 45

Asn His Pro Ser Tyr Ala Arg Gin Gly Gin He Glu Val Ser He Leu 50 55 60 Ala Arg Leu Ser Thr Glu Asn Ala Asp Glu Tyr Asn Phe Val Arg Ala

65 70 75 80

Tyr Glu Thr Phe Gin His Arg Asn His Thr Cys Leu Val Phe Glu Met 85 90 95

Leu Glu Gin Asn Leu Tyr Asp Phe Leu Lys Gin Asn Lys Phe Ser Pro 100 105 110

Leu Pro Leu Lys Val He Arg Pro He Leu Gin Gin Val Ala Thr Ala 115 120 125

Leu Lys Lys Leu Lys Ser Leu Gly Leu He His Ala Asp Leu Lys Pro 130 135 140

Glu Asn He Met Leu Val Asp Pro Val Arg Gin Pro Tyr Arg Val Lys 145 150 155 160

Val He Asp Phe Gly Ser Ala Ser His Val Ser Lys Thr Val Trp Ser

165 170 175

Thr Tyr Leu Gin Ser Arg Tyr Arg Ala Pro Glu He He Leu Gly Leu 180 185 190

Pro Phe Cys Lys Pro He Asp Met Trp Ser Leu Gly Cys Val He Ala 195 200 205

Glu Leu Phe Leu Gly Trp Pro Leu Tyr Pro Gly Ala Leu 210 215 220 (2) INFORMATION FOR SEQ ID NO:7 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 243 amino acids

(B) TYPE: amino acid (C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide (ϋi) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO ( v ) FRAGMENT TYPE : N- terminal

( xi ) SEQUENCE DESCRIPTION : SEQ ID NO : 7 :

Tyr Gin Leu Val Gin His Glu Val Leu Xaa Cys Ser Met Lys Asn Thr

1 5 10 15 Tyr Glu Val Leu Asp Phe Val Gly Arg Gly Thr Phe Gly Gin Val Val

20 25 30

Lys Cys Trp Lys Arg Gly Thr Asn Glu He Val Ala He Lys He Leu 35 40 45

Lys Asn His Pro Ser Tyr Ala Arg Gin Gly Gin He Xaa Xaa Xaa Xaa 50 55 60

Xaa Xaa Glu Val Ser He Leu Ala Arg Leu Ser Xaa Xaa Thr Xaa Glu 65 70 75 80

Asn Ala Asp Glu Tyr Asn Phe Val Xaa Arg Ala Tyr Glu Xaa Thr Xaa

85 90 95 Xaa Phe Gin His Arg Asn His Thr Cys Leu Val Phe Glu Met Leu Glu

100 105 110

Xaa Xaa Gin Asn Leu Tyr Asp Phe Leu Lys Gin Asn Lys Phe Ser Pro 115 120 125

Leu Pro Leu Lys Val He Arg Pro He Leu Gin Gin Val Ala Thr Ala 130 135 140

Leu Lys Lys Leu Lys Ser Leu Gly Leu He His Ala Asp Leu Lys Pro 145 150 155 160

Glu Asn He Met Leu Val Asp Pro Val Arg Gin Pro Tyr Arg Val Lys 165 170 175 Val He Asp Phe Gly Xaa Xaa Ser Ala Ser His Val Ser Xaa Xaa Lys

180 185 190

Thr Val Trp Ser Thr Tyr Xaa Leu Gin Ser Arg Xaa Tyr Arg Ala Pro 195 200 205

Glu He He Leu Gly Leu Pro Phe Cys Lys Pro He Asp Met Trp Ser 210 215 220

Leu Gly Cys Val He Ala Glu Leu Phe Leu Gly Trp Pro Leu Tyr Pro 225 230 235 240

Gly Ala Leu

(2) INFORMATION FOR SEQ ID NO: 8:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 222 amino acids

(B) TYPE: amino acid (C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide (iϋ) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: N-terminal

(Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 8 :

Tyr He Leu Tyr Val Asn Asp Val Leu Gly Val Glu Gin Asn Arg Lys 1 5 10 15 Tyr Leu Val Leu Asp He Leu Gly Gin Gly Thr Phe Gly Gin Val Val

20 25 30

Lys Cys Gin Asn Leu Leu Thr Lys Glu He Leu Ala Val Lys Val Val 35 40 45

Lys Ser Arg Thr Glu Tyr Leu Thr Gin Ser He Thr Glu Ala Lys He 50 55 60

Leu Glu Leu Leu Asn Gin Lys He Asp Pro Thr Asn Lys His His Phe 65 70 75 80

Leu Arg Met Tyr Asp Ser Phe Val His Lys Asn His Leu Cys Leu Val 85 90 95

Phe Glu Leu Leu Ser Asn Asn Leu Tyr Glu Leu Leu Lys Gin Asn Lys 100 105 110

Phe His Gly Leu Ser He Gin Leu He Arg Thr Phe Thr Thr Gin He 115 120 125

Leu Asp Ser Leu Cys Val Leu Lys Glu Ser Lys Leu He His Cys Asp 130 135 140

Leu Lys Pro Glu Asn He Leu Leu Cys Ala Pro Asp Lys Pro Glu Leu 145 150 155 160

Lys He He Asp Phe Gly Ser Ser Cys Glu Glu Ala Arg Thr Val Tyr 165 170 175 Thr Tyr He Gin Ser Arg Phe Tyr Arg Ala Pro Glu He He Leu Gly

180 185 190

He Pro Tyr Ser Thr Ser He Asp Met Trp Ser Leu Gly Cys He Val 195 200 205

Ala Glu Leu Phe Leu Gly He Pro He Phe Pro Gly Ala Ser 210 215 220

(2) INFORMATION FOR SEQ ID NO:9:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 218 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: single (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(v) FRAGMENT TYPE: N-terminal

( i) SEQUENCE DESCRIPTION: SEQ ID NO: 9: Met Ala Ala Thr Arg Tyr Glu Pro Val Ala Glu He Gly Val Gly Ala 1 5 10 15

Tyr Gly Thr Val Tyr Lys Ala Arg Asp Pro His Ser Gly His Phe Val 20 25 30

Ala Leu Lys Ser Val Arg Val Pro Asn Gly Gly Ala Ala Gly Gly Gly

35 40 45 Leu Pro Val Ser Thr Val Arg Glu Val Ala Leu Leu Arg Arg Leu Glu

50 55 60

Ala Phe Glu His Pro Asn Val Val Arg Leu Met Asp Val Cys Ala Thr 65 70 75 80

Ser Arg Thr Asp Arg Asp He Lys Val Thr Leu Val Phe Glu His He 85 90 95

Asp Gin Asp Leu Arg Thr Tyr Leu Asp Lys Ala Pro Pro Pro Gly Leu 100 105 110

Pro Val Glu Thr He Lys Asp Leu Met Arg Gin Phe Leu Ser Gly Leu 115 120 125

Asp Phe Leu His Ala Asn Cys He Val His Arg Asp Leu Lys Pro Glu 130 135 140

Asn He Leu Val Thr Ser Asn Gly Thr Val Lys Leu Ala Asp Phe Gly 145 150 155 160

Leu Ala Arg He Tyr Ser Tyr Gin Met Ala Leu Thr Pro Val Val Val 165 170 175

Thr Leu Trp Tyr Arg Ala Pro Glu Val Leu Leu Gin Ser Thr Tyr Ala 180 185 190

Thr Pro Val Asp Met Trp Ser Val Gly Cys He Phe Ala Glu Met Phe 195 200 205 Arg Arg Lys Pro Leu Phe Cys Gly Asn Ser 210 215

(2) INFORMATION FOR SEQ ID NO:10: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 215 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(v) FRAGMENT TYPE: N-terminal

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:

Tyr Arg Trp Gly Asn Pro Ala Gin Asn Thr Ala Ser Ser Asp Gin Phe 1 5 10 15

Glu Arg Leu Arg Thr Leu Gly Met Gly Ser Phe Gly Arg Val Met Leu 20 25 30 Val Arg His Gin Glu Thr Gly Gly His Tyr Ala Met Lys He Leu Asn 35 40 45

Lys Gin Lys Val Val Lys Met Lys Gin Val Glu His He Leu Asn Glu 50 55 60

Lys Arg He Leu Gin Ala He Asp Phe Pro Phe Leu Val Lys Leu Gin

65 70 75 80 Phe Ser Phe Lys Asp Asn Ser Tyr Leu Tyr Leu Val Met Glu Tyr Val

85 90 95

Pro Gly Gly Glu Met Phe Ser Arg Leu Gin Arg Val Gly Arg Phe Ser 100 105 110

Glu Pro His Ala Cys Phe Tyr Ala Ala Gin Val Val Leu Ala Val Gin

115 120 125

Tyr Leu His Ser Leu Asp Leu He His Arg Asp Leu Lys Pro Glu Asn 130 135 140

Leu Leu He Asp Gin Gin Gly Tyr Leu Gin Val Thr Asp Phe Gly Phe 145 150 155 160

Ala Lys Arg Val Lys Gly Arg Thr Trp Thr Leu Cys Gly Thr Pro Glu 165 170 175

Tyr Leu Ala Pro Glu He He Leu Ser Lys Gly Tyr Asn Lys Ala Val 180 185 190

Asp Trp Trp Ala Leu Gly Val Leu He Tyr Glu Met Ala Val Gly Phe 195 200 205

Pro Pro Phe Tyr Ala Asp Gin 210 215

(2) INFORMATION FOR SEQ ID NO:11:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 219 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(v) FRAGMENT TYPE: N-terminal

(Xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:

Asn Lys Glu Arg Pro Ser Leu Gin He Lys Leu Lys He Glu Asp Phe 1 5 10 15 He Leu His Lys Met Leu Gly Lys Gly Ser Phe Gly Lys Val Phe Leu

20 25 30

Ala Glu Phe Lys Lys Thr Asn Gin Phe Phe Ala He Lys Ala Leu Lys 35 40 45

Lys Asp Val Val Leu Met Asp Asp Asp Val Glu Cys Thr Met Val Glu 50 55 60 Lys Arg Val Leu Ser Leu Ala Trp Glu His Pro Phe Leu Thr His Met 65 70 75 80

Phe Cys Thr Phe Gin Thr Lys Glu Asn Leu Phe Phe Val Met Glu Tyr 85 90 95

Leu Asn Gly Gly Asp Leu Met Tyr His He Gin Ser Cys His Lys Phe 100 105 110 Asp Leu Ser Arg Ala Thr Phe Tyr Ala Ala Glu He He Leu Gly Leu 115 120 125

Gin Phe Leu His Ser Lys Gly He Val Tyr Arg Asp Leu Lys Leu Asp 130 135 140

Asn He Leu Leu Asp Lys Asp Gly His He Lys He Ala Asp Phe Gly 145 150 155 160

Met Cys Lys Glu Asn Met Leu Gly Asp Ala Lys Thr Asn Thr Phe Cys 165 170 175

Gly Thr Pro Asp Tyr He Ala Pro Glu He Leu Leu Gly Gin Lys Tyr 180 185 190 Asn His Ser Val Asp Trp Trp Ser Phe Gly Val Leu Leu Tyr Glu Met 195 200 205

Leu He Gly Gin Ser Pro Phe His Gly Gin Asp 210 215

(2) INFORMATION FOR SEQ ID NO:12:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 3605 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: CDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(Xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:

CAAGTCTTGG TCTACCCACC ATATGTTTAT CAAACTCAGT CAAGTGCCTT TTGTAGTGTG 60

AAGAAACTCA AAGTAGAGCC AAGCAGTTGT GTATTCCAGG AAAGAAACTA TCCACGGACC 120

TATGTGAATG GTAGAAACTT TGGAAATTCT CATCCTCCCA CTAAGGGTAG TGCTTTTCAG 180 ACAAAGATAC CATTTAATAG ACCTCGAGGA CACAACTTTT CATTGCAGAC AAGTGCTGTT 240

GTTTTGAAAA ACACTGCAGG TGCTACAAAG GTCATAGCAG CTCAGGCACA GCAAGCTCAC 300

GTGCAGGCCC CTCAGATTGG GGTGTGGCGA AACAGATTGC ATTTCCTAGA AGGCCCCCAG 360

CGATGTGGAT TGAAGCGCAA GAGTGAGGAG TTGGATAATC ATAGCAGCGC AATGCAGATT 420

GTCGATGAAT TGTCCATACT TCCTGCAATG TTGCAAACCA ACATGGAAAA TCCAGTGACA 480 GTTGTGACAG CTACCACAGG ATCAAAACAG AATTGTACCA CTGGAGAAGG TGACTATCAG 540

TTAGTACAGC ATGAAGTCTT ATGCTCCATG AAAAATACTT ACGAAGTCCT TGATTTTGTT 600 GGTCGAGGCA CGTTTGGCCA GGTAGTTAAA TGCTGGAAAA GAGGGACAAA TGAAATTGTA 660

GCAATCAAAA TTTTGAAGAA TCATCCTTCT TATGCCCGTC AAGGTCAAAT AGAAGTGAGC 720 ATATTAGCAA GGCTCAGTAC TGAAAATGCT GATGAATATA ACTTTGTACG AGCTTATGAA 780

ACGTTTCAGC ACCGTAACCA TACTTGTTTA GTCTTTGAGA TGCTGGAACA AAACTTGTAT 840

GACTTTCTGA AACAAAATAA ATTTAGTCCC CTGCCTCTAA AAGTGATTCG GCCCATTCTT 900

CAACAAGTGG CCACTGCACT GAAAAAATTG AAAAGTCTTG GTTTAATTCA TGCTGATCTA 960

AAGCCTGAGA ATATTATGTT GGTGGATCCT GTTCGGCAGC CTTACAGGGT TAAAGTAATA 1020 GACTTTGGGT CGGCCAGTCA TGTATCAAAG ACTGTTTGGT CAACATATCT ACAATCTCGG 1080

TACAGAGCTC CAGAGATTAT ATTGGGGTTG CCATTTTGTA AGCCAATAGA CATGTGGTCA 1140

TTGGGATGTG TGATTGCAGA ATTATTTCTT GGATGGCCGC TATACCCAGG AGCCTTGGAG 1200

TATGATCAGA TTCGATACAT TTCTCAGACT CAAGGTTTGC CAGGAGAACA GTTGTTAAAT 1260

GTGGGTACTA AATCCACAAG ATTTTTTTGC AAAGAAACAG ATATGTCTCA TTCTGGTTGG 1320 AGATTAAAGA CATTGGAAGA GCATGAGGCA GAGACAGGAA TGAAGTCTAA AGAAGCCAGA 1380

AAATACATTT TCAACAGTCT GGATGATGTA GCGCATGTGA ACACAGTGAT GGATTTGGAA 1440

GGAAGTGATC TTTTTCCTGA GAAAGCTGAT AGAAGAGAAT TTGTTAGTCT GTTGAAGAAA 1500

ATGTTGCTGA TTGATGCAGA TTTAAGAATT ACTCCAGCTG AGACCCTGAA CCATCCTTTT 1560

GTTAATATGA AACATCTTCT AGATTTCCCT CA AGCAACC ATGTAAAGTC CTGTTTTCAT 1620 ATTATGGATA TTTGTAAGTC CCACCTAAAT TCATGTGACA CAAATAATCA CAACAAAACT 1680

TCACTTTTAA GACCAGTTGC TTCAAGCAGT ACTGCTACAC TGACTGCAAA TTTTACTAAA 1740

ATCGGAACAT TAAGAAGTCA GGCATTGACC ACATCTGCTC ATTCAGTTGT GGCGGATGGA 1800

ATACCTCTGC AGGCAGGAAC TCGTCAGTTT GGTTGTGGTG ATGCTTTTCA GCAGACATTG 1860

ATTATCTGTC CCCCAGCTAT TCAAGGTATT CCTGCAACAC ATGGTAAACC CACCAGTTAT 1920 TCAATAAGGG TACATAATAC AGTTCCACTT CTAACTCAGG CCCCAGCTGT GCAGCCACTA 1980

CAGATCCGAC CAAGGAGTTC TTTTCTCAGA CGTCCTCTGG TAGAACACAG ACAGATGCTG 2040

GTGCCTGCCT GGCAACAGGT GACACCCCTG GCTCCTGCTA CTACTACACT AACTTCTGAG 2100

AGTGTGGCTG GTTCACACAG GCTTGGAGAC TGGGGGAAGA TGATTTCATG CAGCAATCAT 2160

TATAACTCAG TGATGCCGCA GCCTCTTCTG ACCAATCAGA TAACTTTATC TGCCCCTCAG 2220 CCAGTTAGTG TGGGGATTGC ACATGTTGTC TGGCCTCAGC CTGCCACTAC CAAGAAAAAT 2280

AAACAGTGCC AGAACAGGAG TAATTCATTA CAGAATACCA ATATCCCACT ATCAGCATTT 2340

ATTTCTCCAA AGATAATTAA TGGGAAAGAT GTCGAGGAAG TAAGTTGTAT AGAAACACAG 2400

GACAATCAGA ACTCAGAAGG AGAGGCAAGA AATTGCTGTG AAACATCTAT CAGACAGGAC 2460

TCTGATTCAT CAGTTTCAGA CAAACAGCGG CAAACCATCA TTATTGCCGA CTCCCCGAGT 2520 CCTGCAGTGA GTGTCATCAC TATCAGCAGT GACACTGATG AGGAAGAGAC TCCCCAGAGA 2580

CATTCACTCA GAGAATGTAA AGGTAGTCTA GATTGTGAAG CTTGCCAGAG CACTTTGAAT 2640 ATTGATCGGA TGTGTTCATT AAGTCCTGAT AGTACTCTGC AGTCCAGCCC ATGCCCCTGC 2700

AAGAGACCGA ATAGTATGTC AGATGAAGAG CAAGAAAGTA GTTGTGATAC GGTGGATGGC 2760 TCTCCGACAT CTGACTCTTC CGGGCATGAC AGTCCATTTG CAGAGAGCAC TTTTGTGGAG 2820

GACACTCATG AAAACACAGA ATTGGTATCC TCTGCTGACA CAGAAACCAA GCCAGCTGTC 2880

TGTTCTGTTG TGGTGCCACC AGTGGAACTA GAAAATGGCT TAAATGCCGA TGAGCATATG 2940

GCAAACACAG ATTCTATATG CCAGCCATTA ATAAAAGGAC GATCTTGCCC CTGGAAGATT 3000

AAACCAGCCT TCTGCAGTGG GTACTCGTCA GCAAAAATTG ACATCAGCAT TCCAGCAGCA 3060 GCATTTGAAC TTCAGTCAGG TTCAGCACTT TGGATCTGGG CTACAAGAGT GGAATGGAAA 3120

CTTTGGGCAC AGAAGACAGC AAGCTTATAT TCCTATCGTG TTACCAGTAA TCCATTCACT 3180

CTTTCTCATG GAAGTCCCAA TCACACAGCA GTGCATGCCC ACCTGGCTGG AAATACACAC 3240

CTCGGAGGAC AGCCTACTCT ACTTCCATAC CCATCATCAG CCACCCTCAG TAGTGCTGCA 3300

CCAGTGGCCC ACCTGTTAGC CTCTCCGTGT ACCTCAAGAC CTATGTTACA GCATCCAACT 3360 TATAATATCT CCCATCCCAG TGGCATACTT CACCAAGTCC CAGTGGGCTT AAATCCCCGT 3420

GTGTTACCAT CCCCAACCAT TCATCAGACT CAGTACAAAC CAATCTTCCC ACCACATTCT 3480

TAACATGCAG CAATCACCTG CATATACTGA TTCCACTGAG TCCAACAAAA CTCAGCCAGT 3540

ATCCATATAT GTGAAAAACA GTATATTGGG AAGCTCAATG ATACAAACAT TTGATTAAAA 3600 ATAAA 3605 (2) INFORMATION FOR SEQ ID NO: 13:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1197 amino acids

(B) TYPE: amino acid (C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide (iϋ) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(v) FRAGMENT TYPE: N-terminal

(Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 13: Gin Val Leu Val Tyr Pro Pro Tyr Val Tyr Gin Thr Gin Ser Ser Ala 1 5 10 15

Phe Cys Ser Val Lys Lys Leu Lys Val Glu Pro Ser Ser Cys Val Phe 20 25 30

Gin Glu Arg Asn Tyr Pro Arg Thr Tyr Val Asn Gly Arg Asn Phe Gly 35 40 45

Asn Ser His Pro Pro Thr Lys Gly Ser Ala Phe Gin Thr Lys He Pro 50 55 60

Phe Asn Arg Pro Arg Gly His Asn Phe Ser Leu Gin Thr Ser Ala Val 65 70 75 80

Val Leu Lys Asn Thr Ala Gly Ala Thr Lys Val He Ala Ala Gin Ala 85 90 95

Gin Gin Ala His Val Gin Ala Pro Gin He Gly Val Trp Arg Asn Arg 100 105 110

Leu His Phe Leu Glu Gly Pro Gin Arg Cys Gly Leu Lys Arg Lys Ser 115 120 125

Glu Glu Leu Asp Asn His Ser Ser Ala Met Gin He Val Asp Glu Leu 130 135 140 Ser He Leu Pro Ala Met Leu Gin Thr Asn Met Glu Asn Pro Val Thr 145 150 155 160

Val Val Thr Ala Thr Thr Gly Ser Lys Gin Asn Cys Thr Thr Gly Glu 165 170 175

Gly Asp Tyr Gin Leu Val Gin His Glu Val Leu Cys Ser Met Lys Asn 180 185 190

Thr Tyr Glu Val Leu Asp Phe Val Gly Arg Gly Thr Phe Gly Gin Val 195 200 205

Val Lys Cys Trp Lys Arg Gly Thr Asn Glu He Val Ala He Lys He

210 215 220 Leu Lys Asn His Pro Ser Tyr Ala Arg Gin Gly Gin He Glu Val Ser

225 230 235 240

He Leu Ala Arg Leu Ser Thr Glu Asn Ala Asp Glu Tyr Asn Phe Val 245 250 255

Arg Ala Tyr Glu Thr Phe Gin His Arg Asn His Thr Cys Leu Val Phe 260 265 270

Glu Met Leu Glu Gin Asn Leu Tyr Asp Phe Leu Lys Gin Asn Lys Phe 275 280 285

Ser Pro Leu Pro Leu Lys Val He Arg Pro He Leu Gin Gin Val Ala 290 295 300 Thr Ala Leu Lys Lys Leu Lys Ser Leu Gly Leu He His Ala Asp Leu 305 310 315 320

Lys Pro Glu Asn He Met Leu Val Asp Pro Val Arg Gin Pro Tyr Arg 325 330 335

Val Lys Val He Asp Phe Gly Ser Ala Ser His Val Ser Lys Thr Val 340 345 350

Trp Ser Thr Tyr Leu Gin Ser Arg Tyr Arg Ala Pro Glu He He Leu 355 360 365

Gly Leu Pro Phe Cys Lys Pro He Asp Met Trp Ser Leu Gly Cys Val 370 375 380 He Ala Glu Leu Phe Leu Gly Trp Pro Leu Tyr Pro Gly Ala Leu Glu 385 390 395 400

Tyr Asp Gin He Arg Tyr He Ser Gin Thr Gin Gly Leu Pro Gly Glu 405 410 415

Gin Leu Leu Asn Val Gly Thr Lys Ser Thr Arg Phe Phe Cys Lys Glu 420 425 430 Thr Asp Met Ser His Ser Gly Trp Arg Leu Lys Thr Leu Glu Glu His 435 440 445

Glu Ala Glu Thr Gly Met Lys Ser Lys Glu Ala Arg Lys Tyr He Phe 450 455 460

Asn Ser Leu Asp Asp Val Ala His Val Asn Thr Val Met Asp Leu Glu 465 470 475 480 Gly Ser Asp Leu Phe Pro Glu Lys Ala Asp Arg Arg Glu Phe Val Ser

485 490 495

Leu Leu Lys Lys Met Leu Leu He Asp Ala Asp Leu Arg He Thr Pro 500 505 510

Ala Glu Thr Leu Asn His Pro Phe Val Asn Met Lys His Leu Leu Asp 515 520 525

Phe Pro His Ser Asn His Val Lys Ser Cys Phe His He Met Asp He 530 535 540

Cys Lys Ser His Leu Asn Ser Cys Asp Thr Asn Asn His Asn Lys Thr 545 550 555 560

Ser Leu Leu Arg Pro Val Ala Ser Ser Ser Thr Ala Thr Leu Thr Ala 565 570 575

Asn Phe Thr Lys He Gly Thr Leu Arg Ser Gin Ala Leu Thr Thr Ser 580 585 590

Ala His Ser Val Val Ala Asp Gly He Pro Leu Gin Ala Gly Thr Arg 595 600 605

Gin Phe Gly Cys Gly Asp Ala Phe Gin Gin Thr Leu He He Cys Pro 610 615 620

Pro Ala He Gin Gly He Pro Ala Thr His Gly Lys Pro Thr Ser Tyr 625 630 635 640 Ser He Arg Val His Asn Thr Val Pro Leu Leu Thr Gin Ala Pro Ala

645 650 655

Val Gin Pro Leu Gin He Arg Pro Arg Ser Ser Phe Leu Arg Arg Pro 660 665 670

Leu Val Glu His Arg Gin Met Leu Val Pro Ala Trp Gin Gin Val Thr 675 680 685

Pro Leu Ala Pro Ala Thr Thr Thr Leu Thr Ser Glu Ser Val Ala Gly 690 695 700

Ser His Arg Leu Gly Asp Trp Gly Lys Met He Ser Cys Ser Asn His

705 710 715 720 Tyr Asn Ser Val Met Pro Gin Pro Leu Leu Thr Asn Gin He Thr Leu

725 730 735

Ser Ala Pro Gin Pro Val Ser Val Gly He Ala His Val Val Trp Pro 740 745 750

Gin Pro Ala Thr Thr Lys Lys Asn Lys Gin Cys Gin Asn Arg Ser Asn 755 760 765

Ser Leu Gin Asn Thr Asn He Pro Leu Ser Ala Phe He Ser Pro Lys 770 775 780

He He Asn Gly Lys Asp Val Glu Glu Val Ser Cys He Glu Thr Gin 785 790 795 800

Asp Asn Gin Asn Ser Glu Gly Glu Ala Arg Asn Cys Cys Glu Thr Ser 805 810 815

He Arg Gin Asp Ser Asp Ser Ser Val Ser Asp Lys Gin Arg Gin Thr 820 825 830

He He He Ala Asp Ser Pro Ser Pro Ala Val Ser Val He Thr He 835 840 845

Ser Ser Asp Thr Asp Glu Glu Glu Thr Pro Gin Arg His Ser Leu Arg 850 855 860 Glu Cys Lys Gly Ser Leu Asp Cys Glu Ala Cys Gin Ser Thr Leu Asn 865 870 875 880

He Asp Arg Met Cys Ser Leu Ser Pro Asp Ser Thr Leu Gin Ser Ser 885 890 895

Pro Cys Pro Cys Lys Arg Pro Asn Ser Met Ser Asp Glu Glu Gin Glu 900 905 910

Ser Ser Cys Asp Thr Val Asp Gly Ser Pro Thr Ser Asp Ser Ser Gly 915 920 925

His Asp Ser Pro Phe Ala Glu Ser Thr Phe Val Glu Asp Thr His Glu

930 935 940 Asn Thr Glu Leu Val Ser Ser Ala Asp Thr Glu Thr Lys Pro Ala Val

945 950 955 960

Cys Ser Val Val Val Pro Pro Val Glu Leu Glu Asn Gly Leu Asn Ala 965 970 975

Asp Glu His Met Ala Asn Thr Asp Ser He Cys Gin Pro Leu He Lys 980 985 990

Gly Arg Ser Cys Pro Trp Lys He Lys Pro Ala Phe Cys Ser Gly Tyr 995 1000 1005

Ser Ser Ala Lys He Asp He Ser He Pro Ala Ala Ala Phe Glu Leu 1010 1015 1020

Gin Ser Gly Ser Ala Leu Trp He Trp Ala Thr Arg Val Glu Trp Lys 1025 1030 1035 1040

Leu Trp Ala Gin Lys Thr Ala Ser Leu Tyr Ser Tyr Arg Val Thr Ser 1045 1050 1055

Asn Pro Phe Thr Leu Ser His Gly Ser Pro Asn His Thr Ala Val His 1060 1065 1070

Ala His Leu Ala Gly Asn Thr His Leu Gly Gly Gin Pro Thr Leu Leu 1075 1080 1085

Pro Tyr Pro Ser Ser Ala Thr Leu Ser Ser Ala Ala Pro Val Ala His 1090 1095 1100 Leu Leu Ala Ser Pro Cys Thr Ser Arg Pro Met Leu Gin His Pro Thr 1105 1110 1115 1120

Tyr Asn He Ser His Pro Ser Gly He Leu His Gin Val Pro Val Gly 1125 1130 1135

Leu Asn Pro Arg Val Leu Pro Ser Pro Thr He His Gin Thr Gin Tyr 1140 1145 1150 Lys Pro He Phe Pro Pro His Ser His Ala Ala He Thr Cys He Tyr

1155 1160 1165

Phe His Val Gin Gin Asn Ser Ala Ser He His He Cys Glu Lys Gin 1170 1175 1180

Tyr He Gly Lys Leu Asn Asp Thr Asn He Leu Lys He

1185 1190 1195

Claims

1. A gene, gene fragment or cDNA coding for any member of a protein kinase family comprising DNA having a partial DNA sequence consisting of the cDNA sequence shown in Table 1, or the sequence given by Sequence ID no. 1, or an equivalent sequence, or a similar sequence having conservatively modified variations to the DNA sequences described by the DNA sequence shown in Table 1, or the sequence given by Sequence ID no. 1, or the cDNA similar to the DNA of the DNA sequence shown in Table 1, or the sequence given by Sequence ID no. 1, except that the cDNA similar to the DNA shown in Table 1, or the sequence given by Sequence ID no. 1, may contain codons coding for any of the amino acids in Table 3, shown as Yakl, Cdk4, PKA, PKC amino acids, that appear in the same column and same position as the amino acid sequence of PKR, but are different than the amino acids of PKR, these codons being substitutes or additions to the codons of the DNA sequence shown in Table 1, such substitutions or additions beginning with the codon that begins at position 535 (TAT) and following in the order shown in Table 3, that allow substitutions or additions to the cDNA sequence in Table 1, or the sequence given by Sequence ID no. 1, that code for amino acids in the same order as are provided by the PKR sequence of Table 3 only including the chosen additions or substitutions.

2. A gene, gene fragment or cDNA, of claim 1, coding for any member of a protein kinase family comprising DNA having a partial DNA sequence consisting of the cDNA sequence shown in Table 1, or the sequence given by Sequence ID no. 1, or an equivalent sequence, or a similar sequence having conservatively modified variations to the DNA sequences described by the DNA sequence shown in Table 1, or the sequence given by Sequence ID no. 1, or the cDNA similar to the DNA of the DNA sequence shown in Table 1, or the sequence given by Sequence ID no. 1, except that the cDNA similar to the DNA shown in Table 1, or the sequence given by

Sequence ID no. 1, may contain codons coding for any of the amino acids in Table 3, shown as the Yakl amino acids, that appear in the same column and same position as the amino acid sequence of PKR, but are different than the amino acids of PKR, these codons being substitutes or additions to the codons of the DNA sequence shown in Table 1, such substitutions or additions beginning with the codon that begins at position 535 (TAT) and following in the order shown in Table 3, that allow substitutions or additions to the cDNA sequence in Table 1, or the sequence given by Sequence ID no. 1, that code for amino acids in the same order as are provided by the PKR sequence of Table 3 only including the chosen additions or substitutions.

3. A gene, gene fragment or cDNA, of claim 2, that codes for a protein having a molecular weight of about 170 kD.

4. A gene, gene fragment or cDNA coding for any member of a protein kinase family comprising a DNA sequence that codes for the peptide sequence shown in

Table 1, or the sequence given by Sequence ID no. 4, or an equivalent sequence, or a similar sequence having conservatively modified variations to the DNA sequences that would code for the sequences consisting of the peptide sequence shown in Table 1, or the sequence given by Sequence ID no. 4, or an equivalent sequence, or a similar sequence having conservatively modified variations to the DNA sequences described by the peptide sequence shown in Table 1, or the sequence of Sequence ID no. 4, or the cDNA coding for the peptide sequence shown in Table 1, or the sequence given by Sequence ID no. 4, except that the cDNA coding for the peptide similar to the peptides shown in Table 1, or the sequence of Sequence ID no. 4, may contain codons coding for any of the amino acids in Table 3, shown as Yakl, Cdk4, PKA, PKC amino acids, that appear in the same column and same position as the amino acid sequence of PKR, but are different than the amino acids of PKR, these codons being substitutes or additions to the codons of the DNA sequence shown in Table 1, such substitutions or additions beginning with the codon that begins at position 535 (TAT) and following in the order shown in Table 3, that allow substitutions or additions to the cDNA sequence in Table 1, or the sequence given by Sequence ID no. 1, that code for amino acids in the same order as are provided by the PKR sequence of Table 3 only including the chosen additions or substitutions.

5. A gene, gene fragment or cDNA of claim 4, coding for any member of a protein kinase family comprising a DNA sequence that codes for the peptide sequence shown in Table 1, or the sequence given by Sequence ID no. 4, or an equivalent sequence, or a similar sequence having conservatively modified variations to the DNA sequences that would code for the sequences consisting of the peptide sequence shown in Table 1, or the sequence given by Sequence ID no. 4, or an equivalent sequence, or a similar sequence having conservatively modified variations to the DNA sequences described by the peptide sequence shown in Table 1, or the sequence of Sequence ID no. 4, or the cDNA coding for the peptide sequence shown in Table 1, or the sequence given by Sequence ID no. 4, except that the cDNA coding for the peptide similar to the peptides shown in Table 1, or the sequence of Sequence ID no. 4, may contain codons coding for any of the amino acids in Table 3, shown as the Yakl amino acids, that appear in the same column and same position as the amino acid sequence of PKR, but are different than the amino acids of PKR, these codons being substitutes or additions to the codons of the DNA sequence shown in Table 1, such substitutions or additions beginning with the codon that begins at position 535 (TAT) and following in the order shown in Table 3, that allow substitutions or additions to the cDNA sequence in Table 1, or the sequence given by Sequence ID no. 1, that code for amino acids in the same order as are provided by the PKR sequence of Table 3 only including the chosen additions or substitutions.

6. A gene, gene fragment or cDNA, of claim 5, that codes for a protein having a molecular weight of about 170 kD.

7. A peptide or protein comprising the proteins having the amino acid sequence shown in Table 1 or shown by Sequence I.D. no. 4, or an equivalent peptide, or a similar peptide having conservatively modified variations, or a peptide comprising the peptide shown in Table 1, or by Sequence ID no. 4 except the equivalent peptide contains any 1 to 243, or any 1 to 20, of the amino acids in Table 3 shown as belonging to Yakl, Cdk4, PKA, or PKC that appear in the same column and same position as the amino acid sequence of PKR, but are different than the amino acids of PKR, these amino acid additions or substitutions begin at position 179 (Tyr) and follow in the same order as shown by Table 3, to make from any 1 to 243, or any 1 to 20, substitutions or additions to the peptide of Table 1 or Sequence ID no. 4.

8. A peptide or protein of claim 7, comprising the proteins having the amino acid sequence shown in Table 1 or shown by Sequence I.D. no. 4, or an equivalent peptide, or a similar peptide having conservatively modified variations, or a peptide comprising the peptide shown in Table 1, or by Sequence ID no. 4 except the equivalent peptide contains any 1 to 243, or any 1-20, of the amino acids in Table 3 shown as belonging to Yakl, that appear in the same column and same position as the amino acid sequence of PKR, but are different than the amino acids of PKR, these amino acid additions or substitutions would begin at position 179 (Tyr) and follow in the same order as shown by Table 3, to make from 1 to 243 substitutions or additions to the peptide of Table 1 or Sequence ID no. 4.

9. A peptide of claim 8, that codes for a protein having a molecular weight of about 170 kD.

10. A gene, gene fragment or cDNA coding for any member of a protein kinase family comprising the sequential amino acids in Table 3, shown as Yakl, Cdk4,

PKA, PKC amino acids, that appear in the same column and same position as the amino acid sequence of PKR, but are different than the amino acids of PKR, these codons being substitutes or additions to the codons of the DNA sequence shown in Table 1, such substitutions or additions beginning with the codon that begins at position 535 (TAT) and following in the order shown in Table 3, that allow substitutions or additions to the cDNA sequence in of PKR, Table 3.

11. A gene, gene fragment or cDNA , of claim 10, coding for any member of a protein kinase family comprising the sequential amino acids in Table 3, shown as the Yakl amino acids, that appear in the same column and same position as the amino acid sequence of PKR, but are different than the amino acids of PKR, these codons being substitutes or additions to the codons of the DNA sequence shown in Table 1, such substitutions or additions beginning with the codon that begins at position 535 (TAT) and following in the order shown in Table 3, that allow substitutions or additions to the cDNA sequence of PKR, Table 3.

12. A peptide or protein comprising the proteins having the amino acid sequence shown in Table 3, shown as PKR, or the amino acid sequence shown in Table 3, that appear in the same column and same position as the amino acid sequence of PKR, but are different than the amino acids of PKR, and that are taken from the Yakl, Cdk4, PKA, PKC amino acids, that appear in the same column and same position as the amino acid sequence of PKR, such substitutions or additions beginning with the first amino acid shown in Table 3, including all possible substitutions or additions to the cDNA sequence in of PKR, Table 3, from all the shown sequences, and continuing to the last amino acid shown in Table 3, including PKR, Yakl, Cdk4, PKA, PKC.

13. A peptide or protein of claim 12, comprising the proteins having the amino acid sequence shown in Table 3, shown as PKR, or the amino acid sequence shown in Table 3, that appear in the same column and same position as the amino acid sequence of PKR, but are different than the amino acids of PKR, and that are taken from the Yakl amino acids, that appear in the same column and same position as the amino acid sequence of PKR, such substitutions or additions beginning with the first amino acid shown in Table 3, including all possible substitutions or additions to the cDNA sequence in PKR, Table 3, from all the shown sequences, and continuing to the last amino acid shown in Table 3, including PKR and Yakl.