Genome instability of mis-match repair and its role in carcinogenesis due to radiation

Homologue genes, mutS and mutL have been known as a key gene for mismatch repair in E. coli. In this study, identification of such homologue genes in the nematode was attempted using caenorhabditis elegans and three kinds of mutS homologues (mshG, mshZ and mshF) and 2 kinds of mutL ones were identified. From after isolation of these genes, base sequences were analyzed. Then, an insertion mutant in which Tcl transposon is inserted in Exon 13 positioned at the center of mshF was screened and its homozygote where breakage of transposon in somatic cells occurred frequently was obtained and its morphological changes were not significant. In the nematoda, we detected a highly conserved domain in mutS family gene, which is commonly present in yeast and human genes. Based on the amino acid sequence of this domain, four kinds of primers were constructed for PCR reaction using the whole DNA from the nematoda as a template and four DNA fragments of which sizes were almost corresponding to the homologue proteins were produced. From screening of Tc1 insertion mutant for 8 mismatch repair genes, three strains; mshF, rqhW and RqhY were obtained and the gene structures and the positions of Tc1 insertion in these strains were determined. The sensitivities to ionizing radiation, UV and alkyl reagent of these strains were not significantly different from those of the wild strain. (M.N.)