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Missense variants in hMLH1 identified in patients from the German HNPCC consortium and functional studies

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Abstract

Missense mutations of the DNA mismatch repair gene MLH1 are found in a significant fraction of patients with Lynch syndrome (hereditary nonpolyposis colorectal cancer, HNPCC) and their pathogenicity often remains unclear. We report here all 88 MLH1 missense variants identified in families from the German HNPCC consortium with clinical details of these patients/families. We investigated 23 MLH1 missense variants by two functional in vivo assays in yeast; seven map to the ATPase and 16 to the protein interaction domain. In the yeast-2-hybrid (Y2H) assay three variants in the ATPase and twelve variants in the interaction domain showed no or a reduced interaction with PMS2; seven showed a normal and one a significantly higher interaction. Using the Lys2A 14 reporter system to study the dominant negative mutator effect (DNE), 16 variants showed no or a low mutator effect, suggesting that these are nonfunctional, three were intermediate and four wild type in this assay. The DNE and Y2H results were concordant for all variants in the interaction domain, whereas slightly divergent results were obtained for variants in the ATPase domain. Analysis of the stability of the missense proteins in yeast and human embryonic kidney cells (293T) revealed a very low expression for seven of the variants in yeast and for nine in human cells. In total 15 variants were classified as deleterious, five were classified as variants of unclassified significance (VUS) and three were basically normal in the functional assays, P603R, K618R, Q689R, suggesting that these are neutral.

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Acknowledgments

This work was supported by grants from the German Cancer Aid (Deutsche Krebshilfe e.V. Bonn, project nos 70-2397, 108132 and 108628) and a multicentre grant from the German Cancer Aid (project nos. 70-2371, 106244 and 107318).

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Authors and Affiliations

  1. Institute of Human Genetics and Anthropology, Heinrich Heine University and University Hospital Duesseldorf, Postfach 101007, 40001, Duesseldorf, Germany

    Karin Hardt, Beate Betz, Timm Goecke, Haniyeh Yazdanparast, Robin Küppers, Kati Servan & Brigitte Royer-Pokora

  2. Institute of Functional Microbial Genomics, Heinrich Heine University, Duesseldorf, Germany

    Sven Boris Heick & Johannes H. Hegemann

  3. Institute of Human Genetics, University Hospital Bonn, Bonn, Germany

    Verena Steinke & Nils Rahner

  4. Center of Medical Genetics, University Hospital of the Ludwig-Maximilians-University Munich and MGZ, Munich, Germany

    Monika Morak & Elke Holinski-Feder

  5. Institute of Medical Informatics, Statistics and Epidemiology, University Leipzig, Leipzig, Germany

    Christoph Engel

  6. Department of Surgery, HELIOS St. Josefs-Hospital, Bochum, Germany

    Gabriela Möslein

  7. Department of Surgical Research, University of Technology Dresden, Dresden, Germany

    Hans-Konrad Schackert

  8. Institute of Applied Tumor Biology, University Hospital Heidelberg, Heidelberg, Germany

    Magnus von Knebel Doeberitz

  9. Department of Medicine, Knappschaftskrankenhaus, Ruhr-University Bochum, Bochum, Germany

    Christian Pox

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  1. Karin Hardt
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  2. Sven Boris Heick
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  14. Hans-Konrad Schackert
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  15. Magnus von Knebel Doeberitz
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  16. Christian Pox
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  17. Johannes H. Hegemann
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  18. Brigitte Royer-Pokora
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Peter Propping and the German HNPCC consortium

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Correspondence to Brigitte Royer-Pokora.

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Hardt, K., Heick, S.B., Betz, B. et al. Missense variants in hMLH1 identified in patients from the German HNPCC consortium and functional studies. Familial Cancer 10, 273–284 (2011). https://doi.org/10.1007/s10689-011-9431-4

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  • DOI: https://doi.org/10.1007/s10689-011-9431-4

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