Functional analysis of human MLH1 mutations in Saccharomyces cerevisiae (original) (raw)

Nature Genetics volume 19, pages 384–389 (1998)Cite this article

A Correction to this article was published on 01 February 1999

Abstract

Hereditary non-polyposis colorectal cancer (HNPCC; OMIM 120435-6) is a cancer-susceptibility syndrome1 linked to inherited defects in human mismatch repair (MMR) genes2. 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.

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Acknowledgements

We are grateful to the patients who participated in this study. We thank T. D. Petes for providing the pSH31 plasmid, N. Papadopoulos and S. Mizutani for providing cDNA containing hMLH1 mutations and E. O'Neil for technical assistance. We also thank A. Yasui, T. Noda, M. Vidal, K. Umezu, T. Hunato, S. Ishii, S. Kure and A. Horii for helpful comments about this work. This work was supported in part by Grant-in Aids for Scientific Research and International Scientific Research (Joint Study) from the Ministry of Education, Science, Sports and Culture (C.I., S.H.F., R.D.K.), Grant-in Aid from the Ministry of Health and Welfare (C.I.), the Osaka Cancer Research Foundation (C.I.), the Tokyo Biochemical Research Foundation (C.I.), the Sapporo Bioscience Foundation (C.I.) and a National Institutes of Health grant, no. GM50006 (R.D.K.).

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Author notes

  1. Richard D. Kolodner
    Present address: Ludwig Institute for Cancer Research, University of California San Diego School of Medicine, La Jolla, California, 92093, USA

Authors and Affiliations

  1. Department of Clinical Oncology, Institute of Development, Aging and Cancer, Tohoku University, Sendai, 980-8575, Japan
    Hideki Shimodaira, Hiroyuki Shibata, Takao Suzuki, Ryunosuke Kanamaru & Chikashi Ishioka
  2. Charles A. Dana Division of Human Cancer Genetics, Dana-Farber Cancer Institute, Boston, 02115, Massachusetts, USA
    Nicole Filosi & Richard D. Kolodner
  3. Division of Experimental Oncology, Istituto Nazionale Tumori, Milano, 20133, Italy
    Paolo Radice
  4. The Seattle Project, Fred Hutchinson Cancer Research Center, Seattle, 98109, Washington, USA
    Stephen H. Friend

Authors

  1. Hideki Shimodaira
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  2. Nicole Filosi
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  3. Hiroyuki Shibata
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  4. Takao Suzuki
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  5. Paolo Radice
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  6. Ryunosuke Kanamaru
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  7. Stephen H. Friend
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  8. Richard D. Kolodner
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  9. Chikashi Ishioka
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Corresponding authors

Correspondence toRichard D. Kolodner or Chikashi Ishioka.

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Shimodaira, H., Filosi, N., Shibata, H. et al. Functional analysis of human MLH1 mutations in Saccharomyces cerevisiae.Nat Genet 19, 384–389 (1998). https://doi.org/10.1038/1277

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