Functional analysis of human MLH1 mutations in Saccharomyces cerevisiae

Hideki Shimodaira, Nicole Filosi, Hiroyuki Shibata, Takao Suzuki, Paolo Radice, Ryunosuke Kanamaru, Stephen H. Friend, Richard D. Kolodner, Chikashi Ishioka

研究成果: Article査読

120 被引用数 (Scopus)

抄録

Hereditary non-polyposis colorectal cancer (HNPCC; OMIM 120435-6) is a cancer-susceptibility syndrome linked to inherited defects in human mismatch repair (MMR) genes. Germline missense human MLH1 (hMLH1) mutations are frequently detected in HNPCC (ref. 3), making functional characterization of mutations in hMLH1 critical to the development of genetic testing for HNPCC. Here, we describe a new method for detecting mutations in hMLH1 using a dominant mutator effect of hMLH1 cDNA expressed in Saccharomyces cerevisiae. The majority of hMLH1 missense mutations identified in HNPCC patients abolish the dominant mutator effect. Furthermore, PCR amplification of hMLH1 cDNA from mRNA from a HNPCC patient, followed by in vivo recombination into a gap expression vector, allowed detection of a heterozygous loss-of-function missense mutation in hMLH1 using this method. This functional assay offers a simple method for detecting and evaluating pathogenic mutations in hMLH1.

本文言語English
ページ(範囲)384-389
ページ数6
ジャーナルNature Genetics
19
4
DOI
出版ステータスPublished - 1998

ASJC Scopus subject areas

  • Genetics

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