Vertebrate telomerase genes and proteins and uses thereof

Abstract

Nucleic acid molecules encoding vertebrate telomerase are provided. Gene products, expression vectors and host cells suitable for expressing telomerase are also provided. Methods for identifying inhibitors of telomerase activity and inhibitor compositions are disclosed.

1 . An isolated nucleic acid molecule encoding a splice variant of a reference human telomerase, wherein the reference human telomerase has the amino acid sequence of SEQ ID NO: 2.  
     
     
         2 . An isolated nucleic acid molecule, comprising any of SEQ ID NOS: 34, 36, 38, 41, 43, 45, 47, or 49 or a hybridizes under normal stringency conditions to the complement of the sequences thereof, wherein the nucleic acid molecule is not SEQ ID NO:1 or encodes SEQ ID NO:2.  
     
     
         3 . An isolated nucleic acid molecule comprising any of SEQ ID NO: 23, 25, 27, 29, 30, 32, or 33, or hybridizes under normal stringency conditions to the complement of the sequences thereof, wherein the nucleic acid molecule is not SEQ ID NO:1 or encodes SEQ ID NO:2.  
     
     
         4 . An oligonucleotide comprising from 10 to 100 contiguous nucleotides of SEQ ID NO: 23, 25, 27, 29, 30, 32, or 33 or of the complements thereof or has 90% sequence identity to one of the sequences or complements.  
     
     
         5 . The oligonucleotide of  claim 4 , wherein the oligonucleotide is labeled.  
     
     
         6 . The oligonucleotide of  claim 5 , wherein the label is a radiolabel, a chemiluminescent label, or biotin.  
     
     
         7 . An expression vector, comprising a heterologous promoter operably linked to a nucleic acid molecule according any of claims  1 - 3 .  
     
     
         8 . The expression vector of  claim 7 , wherein the vector is selected from the group consisting of bacterial vectors, retroviral vectors, adenoviral vectors and yeast vectors.  
     
     
         9 . A host cell containing a vector according to  claim 7 .  
     
     
         10 . The host cell of  claim 9 , wherein the cell is selected from the group consisting of human cell, monkey cell, mouse cell, rat cell, yeast cell and bacterial cell.  
     
     
         11 . An isolated protein comprising a splice variant of a reference human telomerase protein, wherein the reference protein has the amino acid sequence of SEQ ID NO: 2.  
     
     
         12 . The protein of  claim 11 , wherein the protein comprises one SEQ ID NOS: 35, 37, 39, 42, 44, 46, 48, or 50, wherein the protein is not SEQ ID NO: 2.  
     
     
         13 . A peptide having one of SEQ ID NOS: 24, 26, 28, or 31 or has a sequence 90% identical to one of the sequences.  
     
     
         14 . A nucleic acid probe that is capable of specifically hybridizing to SEQ ID NOS: 23, 29, 30, 32 or 33.  
     
     
         15 . The probe of  claim 14 , wherein the probe is from 12 to 200 nucleotides long.  
     
     
         16 . The probe of  claim 14 , wherein the nucleic acid molecule is labeled.  
     
     
         17 . A pair of oligonucleotide primers capable of specifically amplifying all or a portion of a nucleic acid molecule encoding a splice variant of a reference human telomerase, wherein the reference telomerase has SEQ ID NO:1.  
     
     
         18 . The primers of  claim 17 , wherein the splice variant has the sequence of any of SEQ ID NOS: 23, 25, 27, 29, 30, 32, or 33.  
     
     
         19 . The primers of  claim 17 , wherein the primers flank nucleotide 222, 1950, 2131-2166, 2287-2468, 2843, or 3157 of SEQ ID NO:1.  
     
     
         20 . The primers of  claim 17 , wherein only one of each primer pair flanks nucleotide 222, 1950, 2131-2166, 2287-2468, 2843, or 3157 of SEQ ID NO:1 and the other primer of the pair has sequence derived from one SEQ ID NOS: 23, 25, 27, 29, 30, 32, or 33 or complements thereof.  
     
     
         21 . A method of diagnosing cancer in a patient, comprising preparing tumor cDNA and amplifying the tumor cDNA using primers that specifically amplify a splice variant of a reference human telomerase nucleic acid sequence, wherein the detection of a splice variant telomerase nucleic acid sequence is indicative of a diagnosis of cancer.  
     
     
         22 . The method of  claim 21 , wherein the primers flank nucleotide 222, 1950, 2131-2166, 2287-2468, 2843, or 3157 of SEQ ID NO:1. or wherein one of each primer pair flanks nucleotide 222, 1950, 2131-2166, 2287-2468, 2843, or 3157 of SEQ ID NO:1 and the other primer of the pair has sequence derived from one SEQ ID NOS: 23, 25, 27, 29, 30, 32, or 33 or complements thereof.  
     
     
         23 . A method of determining a pattern of telomerase RNA expression in cells, comprising preparing cDNA from mRNA isolated from the cells, amplifying the cDNA using primers that amplify a reference human telomerase sequence and one or more splice variant telomerase sequences, therefrom determining the pattern of telomerase RNA expression.  
     
     
         24 . The method of  claim 23 , further comprising detecting the amplified product by hybridization with one or more oligonucleotides having all or part of SEQ ID NO: 23, 25, 27, 29, 30, 32, or 33 or the complements thereof.  
     
     
         25 . A method of diagnosing cancer in a patient, comprising determining a pattern of telomerase RNA expression according to  claim 23 , therefrom determining the pattern of telomerase RNA expression, wherein the pattern is indicative of a diagnosis of cancer.  
     
     
         26 . The method of  claim 25 , further comprising comparing the pattern to a pattern obtained from normal cells.  
     
     
         27 . A non-human transgenic animal whose cells contain a splice variant telomerase sequence that is operably linked to a promoter effective for the expression of the gene.  
     
     
         28 . A method of identifying an effector of telomerase activity comprising: 
 (a) adding a candidate effector to a mixture of a splice variant of a human telomerase protein, RNA component and template;    (b) detecting telomerase activity; and    (c) comparing the amount of activity in step (b) to the amount of activity in a control mixture without candidate effector, therefrom identifying an effector.