A yeast two-hybrid assay provides a simple way to evaluate the vast majority of hMLH1 germ-line mutations

Emiko Kondo, Hideyuki Suzuki, Akira Horii, Shinichi Fukushige

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61 Citations (Scopus)

Abstract

Germ-line mutations in the hMLH1 gene are the most frequent cause of hereditary nonpolyposis colorectal cancer and are characterized by mis-sense mutations at high frequency. We found a yeast two-hybrid assay to be an extremely useful and simple tool for evaluating the biological significance of such hMLH1 germ-line missense mutations; 78% (18 of 23) of the missense ones can be recognized as causative for nonpolyposis colorectal cancer. In addition, two of five variants not recognized as causative were thought to he rare polymorphisms. However, we could not detect any differences between wild-type hMLH1 and any of the nine already known polymorphisms causing amino acid alterations. Additional analysis demonstrated that the two-hybrid assay not only detected the dysfunctions at the COOH terminus of the hMLH1 protein necessary for the interaction with associated proteins but also detected a conformational change at the NH2 terminus carrying ATPase activity. Thus, this method provides a simple and reliable system for accurate diagnosis of hMLH1 alterations.

Original languageEnglish
Pages (from-to)3302-3308
Number of pages7
JournalCancer Research
Volume63
Issue number12
Publication statusPublished - 2003 Jun 15

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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