Method of Detection of Alterations in MSH5

Abstract

We have now discovered that mammals have a DNA gene analogous to that existing in bacteria. MSH5 defects or alterations in this mismatch repair pathway in a mammal, such as a human can be diagnostic of a predisposition to cancer, and prognostic for a particular cancer. We have discovered and sequenced MSH5 in this in a number of mammals, including humans. This gene can be used in assays, to express gene product, for drug screens, and therapeutically.

1 . An isolated and purified human MSH5 protein having the amino acid sequence set forth in SEQ ID NO:2, or a fragment of at least six amino acids thereof. 
     
     
         2 .- 12 . (canceled) 
     
     
         13 . A method of determining whether there is an alteration in a mammalian MSH5 gene which comprises: a) isolating a biological specimen from a preselected mammal; b) testing the specimen for an alteration in said mammalian MSH5 protein and c) comparing the results obtained in step b) with a wild type control. 
     
     
         14 . The method of  claim 13 , wherein the biological specimen is selected from blood, tissue, serum, stool, urine, sputum, cerebrospinal fluid, supernatant from cell lysate and a eukaryotic cell sample. 
     
     
         15 . The method of  claim 13 , wherein the mammal is a human. 
     
     
         16 . The method of  claim 13 , wherein an alteration is indicative of a predisposition to malignant growth of cells in the mammal. 
     
     
         17 . The method of  claim 13 , wherein an alteration is indicative of a predisposition to a malady associated with inappropriate meiotic segregation. 
     
     
         18 . The method of  claim 15 , wherein the biological specimen is selected from a group of blood related individuals. 
     
     
         19 . (canceled) 
     
     
         20 . The method of  claim 17 , wherein the malady is infertility or Downs Syndrome. 
     
     
         21 .- 27 . (canceled) 
     
     
         28 . The method of  claim 13 , wherein the alteration is detected by measuring the level of protein expression. 
     
     
         29 .- 32 . (canceled) 
     
     
         33 . A method of diagnosing a DNA mismatch repair defective tumor of a mammal, comprising: isolating a tissue from said mammal suspected of being a tumor;
 and detecting an alteration in a MSH5 expression product, wherein said alteration is indicative of a DNA mismatch repair defective tumor.   
     
     
         34 . The method of  claim 33 , wherein the mammal is a human. 
     
     
         35 . The method of  claim 34 , wherein the DNA mismatch repair defective tumor is lung, breast, colorectal ovary, endometrial (uterine), renal, bladder, skin, rectal and small bowel. 
     
     
         36 . A method of prognosis in an individual having cancer, comprising, comparing a cancer cell from said individual with a non-cancer cell from said individual for the presence of an alteration in the MSH5 protein. 
     
     
         37 . The method of  claim 36 , wherein an alteration in both cells indicates a genetic basis for said cancer. 
     
     
         38 . A method of screening for agents affecting a mammalian MSH5 protein comprising:
 a) selecting a first test cell having an alteration in the mammalian MSH5 protein;   b) selecting a second test cell, said second cell derived from said first cell, but not having the alteration in the MSH5 protein;   c) contacting said test cells with a selected agent; and   d) comparing the effects of said agent on the first and second test cells.   
     
     
         39 . An isolated polypeptide encoded by an isolated nucleic acid encoding at least one exon of human MSH5 or a fragment encoded by at least 25 nucleotides of an exon from SEQ ID NO: 1. 
     
     
         40 . An isolated polypeptide segment, wherein said polypeptide segment is encoded by at least 25 nucleotides between nucleotides 235-1908 of SEQ ID NO: 1. 
     
     
         41 . An isolated polypeptide segment encoded by at least 25 nucleotides, from at least one exon from SEQ ID NO: 1, wherein the nucleotide sequence is selected from the group consisting of exons 1-24, wherein exon 1 starts at position 235 and continues continuously for 221 basepairs, exon 2 is the next 160 basepairs, exon 3 is the next 124 basepairs, exon 4 is the next 81 basepairs, exon 5 is the next 63 basepairs, exon 6 is the next 122 basepairs, exon 7 is the next 110 basepairs, exon 8 is the next 36 basepairs, exon 9 is the next 83 basepairs, exon 10 is the next 46 basepairs, exon 11 is the next 139 basepairs, exon 12 is the next 63 basepairs, exon 13 is the next 129 basepairs, exon 14 is the next 73 basepairs, exon 15 is the next 110 basepairs, exon 16 is the next 81 basepairs, exon 17 is the next 88 basepairs, exon 18 is the next 190 basepairs, exon 19 is the next 127 basepairs, exon 20 is the next 150 basepairs, exon 21 is the next 75 basepairs, exon 22 is the next 144 basepairs, exon 23 is the next 138 basepairs and exon 24 is the next 74 basepairs.