Human Prt1-like subunit protein (hPrt1) and human eIF4G-like protein (p97) genes

Abstract

The present invention relates to novel human Prt1 (hPrt1) and eIF4G-like (p97) proteins which are involved in eukaryotic transcription In particular, isolated nucleic acid molecules are provided encoding the human hPrt1 and p97 proteins. hPrt1 and p97 polypeptides are also provided, as are vectors, host cells and recombinant methods for producing the same. The invention further relates to screening methods for identifying agonists and antagonists of hPrt1 and p97 activity. Also provided are therapeutic methods for treating disease states associated with the hPrt1 and p97 proteins.

What is claimed is:  
     
         1 . An isolated nucleic acid molecule comprising a polynucleotide having a nucleotide sequence at least 95% identical to a sequence selected from the group consisting of: 
 (a) a nucleotide sequence encoding the hPrt1 polypeptide having the complete amino acid sequence in SEQ ID NO:2;    (b) a nucleotide sequence encoding the hPrt1 polypeptide having the amino acid sequence at positions from about 2 to about 873 in SEQ ID NO:2;    (c) a nucleotide sequence encoding the hPrt1 polypeptide having the complete amino acid sequence encoded by the cDNA clone contained in ATCC Deposit Number 97766;    (d) a nucleotide sequence encoding the p97 polypeptide having the complete amino acid sequence in SEQ ID NO:4;    (e) a nucleotide sequence encoding the p97 polypeptide having the amino acid sequence at positions from about 2 to about 907 in SEQ ID NO:4;    (f) a nucleotide sequence encoding the p97 polypeptide having the complete amino acid sequence encoded by the cDNA clone contained in ATCC Deposit Number 97767; and    (g) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b), (c), (d), (e) or (f).    
     
     
         2 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the complete nucleotide sequence in SEQ ID NO: 1 or SEQ ID NO:3.  
     
     
         3 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has either the nucleotide sequence in SEQ ID NO: 1 encoding a polypeptide having the complete amino acid sequence in SEQ ID NO:2 or the nucleotide sequence in SEQ ID NO:3 encoding a polypeptide having the complete amino acid sequence in SEQ ID NO:4.  
     
     
         4 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the complete nucleotide sequence of the cDNA clones contained in ATCC Deposit Number 97766 or ATCC Deposit Number 97767.  
     
     
         5 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence encoding the polypeptides having the complete amino acid sequence encoded by the CDNA clones contained in ATCC Deposit Number 97766 or ATCC Deposit Number 97767.  
     
     
         6 . An isolated nucleic acid molecule comprising a polynucleotide which hybridizes under stringent hybridization conditions to a polynucleotide having a nucleotide sequence identical to a nucleotide sequence in (a), (b), (c), (d), (e), (f) or (g) of  claim 1  wherein said polynucleotide which hybridizes does not hybridize under stringent hybridization conditions to a polynucleotide having a nucleotide sequence consisting of only A residues or of only T residues.  
     
     
         7 . An isolated nucleic acid molecule comprising a polynucleotide which encodes the amino acid sequence of an epitope-bearing portion of a polypeptide having an amino acid sequence in (a), (b), (c), (d), (e) or (f) of  claim 1 .  
     
     
         8 . The isolated nucleic acid molecule of  claim 7 , which encodes an epitope-bearing portion of an hPrt1 polypeptide selected from the group consisting of: a polypeptide comprising amino acid residues from about 1 to about 188 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 193 to about 235 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 248 to about 262 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 270 to about 350 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 361 to about 449 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 458 to about 620 in SEQ ID NO:2; and a polypeptide comprising amino acid residues from about 639 to about 846 in SEQ ID NO:2 or an epitope-bearing portion of a p97 polypeptide selected from the group consisting of: a polypeptide comprising amino acid residues from about 1 to about 98 in SEQ ID NO:4; a polypeptide comprising amino acid residues from about 121 to about 207 in SEQ ID NO:4; a polypeptide comprising amino acid residues from about 232 to about 278 in SEQ ID NO:4; a polypeptide comprising amino acid residues from about 287 to about 338 in SEQ ID NO:4; a polypeptide comprising amino acid residues from about 347 to about 578 in SEQ ID NO:4; a polypeptide comprising amino acid residues from about 593 to about 639 in SEQ ID NO:4; a polypeptide comprising amino acid residues from about 681 to about 770 in SEQ ID NO:4; a polypeptide comprising amino acid residues from about 782 to about 810 in SEQ ID NO:4; and a polypeptide comprising amino acid residues from about 873 to about 905 in SEQ ID NO:4.  
     
     
         9 . A method for making a recombinant vector comprising inserting an isolated nucleic acid molecule of  claim 1  into a vector.  
     
     
         10 . A recombinant vector produced by the method of  claim 9 .  
     
     
         11 . A method of making a recombinant host cell comprising introducing the recombinant vector of  claim 10  into a host cell.  
     
     
         12 . A recombinant host cell produced by the method of  claim 11 .  
     
     
         13 . A recombinant method for producing a polypeptide, comprising culturing the recombinant host cell of  claim 12  under conditions such that said polypeptide is expressed and recovering said polypeptide.  
     
     
         14 . An isolated polypeptide having an amino acid sequence at least 95% identical to a sequence selected from the group consisting of: 
 (a) the amino acid sequence of the hPrt1 polypeptide having the complete amino acid sequence in SEQ ID NO:2;    (b) the amino acid sequence of the hPrt1 polypeptide having the amino acid sequence at positions from about 2 to about 873 in SEQ If) NO:2;    (c) the amino acid sequence of the hPrt1 polypeptide having the complete amino acid sequence encoded by the cDNA clone contained in ATCC Deposit Number 97766;    (d) the amino acid sequence of the p97 polypeptide having the complete amino acid sequence in SEQ ID NO:4;    (e) the amino acid sequence of the p97 polypeptide having the amino acid sequence at positions from about 2 to about 907 in SEQ ID NO:4;    (f) the amino acid sequence of the p97 polypeptide having the complete amino acid sequence encoded by the cDNA clone contained in ATCC Deposit Number 97767; and    (g) the amino acid sequence of an epitope-bearing portion of any one of the polypeptides of (a), (b), (c), (d), (e) or (f).    
     
     
         15 . An isolated polypeptide comprising an epitope-bearing portion of either the hPrt1 protein, wherein said portion is selected from the group consisting of: a polypeptide comprising amino acid residues from about 1 to about 188 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 193 to about 235 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 248 to about 262 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 270 to about 350 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 361 to about 449 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 458 to about 620 in SEQ ID NO:2; and a polypeptide comprising amino acid residues from about 639 to about 846 in SEQ ID NO:2 or the p97 protein, wherein said portion is selected from the group consisting of: a polypeptide comprising amino acid residues from about 1 to about 98 in SEQ ID NO:4; a polypeptide comprising amino acid residues from about 121 to about 207 in SEQ ID NO:4; a polypeptide comprising amino acid residues from about 232 to about 278 in SEQ ID NO:4; a polypeptide comprising amino acid residues from about 287 to about 338 in SEQ ID NO:4; a polypeptide comprising amino acid residues from about 347 to about 578 in SEQ ID NO:4; a polypeptide comprising amino acid residues from about 593 to about 639 in SEQ ID NO:4; a polypeptide comprising amino acid residues from about 681 to about 770 in SEQ ID NO:4; a polypeptide comprising amino acid residues from about 782 to about 810 in SEQ ID NO:4; and a polypeptide comprising amino acid residues from about 873 to about 905 in SEQ ID NO:4.  
     
     
         16 . An isolated antibody that binds specifically to a polypeptide of  claim 15 .  
     
     
         17 . A method of translating mRNA in vitro, comprising: 
 (a) a translation reaction mixture containing both labeled and unlabeled amino acids;    (b) mixing an effective amount of hPrt1 and/or p97 protein;    (c) fractionating the sample to separate labeled protein; and    (d) detecting the presence of the labeled protein.    
     
     
         18 . A method of screening for agonists and antagonists of hPrt1 and/or p97 activity comprising: 
 (a) contacting cells which express hPrt1 and/or p97 polypeptides with a candidate compound,    (b) assaying a cellular response, and    (c) comparing the cellular response to a standard cellular response made in absence of the candidate compound;    whereby, an increased cellular response over the standard indicates that the compound is an agonist and a decreased cellular response over the standard indicates that the compound is an antagonist.    
     
     
         19 . A method of treating a disease state associated with apoptosis comprising introducing an effective amount of an hPrt1 and/or p97 protein into an individual to be treated in admixture with a pharmaceutically acceptable carrier.