Functional, structural, and genetic evaluation of 20 CDKN2A germ line mutations identified in melanoma-prone families or patients

Authors

  • Caroline Kannengiesser,

    1. Service de Génétique, Institut Gustave Roussy, Villejuif, France
    2. Laboratoire “Génomes et cancers,” FRE2939 CNRS, Institut Gustave Roussy, Université Paris-Sud, Orsay, France
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  • Sharon Brookes,

    1. Cancer Research UK, London Research Institute, London, United Kingdom
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  • Anna Gutierrez del Arroyo,

    1. Cancer Research UK, London Research Institute, London, United Kingdom
    Current affiliation:
    1. Department of Medicine and Wolfson Institute of Biomedical Research, University College London, Jules Thorn Building, Middlesex Hospital Mortimer St, London W1T 3AA, United Kingdom
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  • Danielle Pham,

    1. Service de Génétique, Institut Gustave Roussy, Villejuif, France
    2. Laboratoire “Génomes et cancers,” FRE2939 CNRS, Institut Gustave Roussy, Université Paris-Sud, Orsay, France
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  • Johny Bombled,

    1. Service de Génétique, Institut Gustave Roussy, Villejuif, France
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  • Michel Barrois,

    1. Service de Génétique, Institut Gustave Roussy, Villejuif, France
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  • Olivier Mauffret,

    1. Département de Biologie et Pharmacologie Structurales, Unité Mixte de Recherche (UMR) 8113 Centre National de la Recherche Scientifique (CNRS)-LBPA (ENS Cachan), Institut Gustave Roussy, Villejuif, France
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  • Marie-Françoise Avril M,

    1. Département de médecine, Institut Gustave Roussy, Villejuif, France
    Current affiliation:
    1. AP–HP Service de Dermatologie, Hôpital Cochin, Pavillon Tarnier, 89, rue d'Assas, 75006 Paris, France
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  • Agnès Chompret,

    1. Département de médecine, Institut Gustave Roussy, Villejuif, France
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    • The members French Hereditary Melanoma Study Group are listed in the Acknowledgments section.

  • Gilbert M. Lenoir,

    1. Service de Génétique, Institut Gustave Roussy, Villejuif, France
    2. Laboratoire “Génomes et cancers,” FRE2939 CNRS, Institut Gustave Roussy, Université Paris-Sud, Orsay, France
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  • Alain Sarasin,

    1. Laboratoire “Génomes et cancers,” FRE2939 CNRS, Institut Gustave Roussy, Université Paris-Sud, Orsay, France
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  • Gordon Peters,

    1. Cancer Research UK, London Research Institute, London, United Kingdom
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  • Brigitte Bressac-de Paillerets

    Corresponding author
    1. Service de Génétique, Institut Gustave Roussy, Villejuif, France
    2. Laboratoire “Génomes et cancers,” FRE2939 CNRS, Institut Gustave Roussy, Université Paris-Sud, Orsay, France
    • Service de Génétique, Institut Gustave Roussy, 39 rue Camille Desmoulins, 94805, Villejuif, France
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  • Communicated by David E. Goldgar

  • This work is dedicated to our late colleague Agnès Chompret.

Abstract

Germline mutations of the CDKN2A gene are found in melanoma-prone families and individuals with multiple sporadic melanomas. The encoded protein, p16INK4A, comprises four ankyrin-type repeats, and the mutations, most of which are missense and occur throughout the entire coding region, can disrupt the conformation of these structural motifs as well as the association of p16INK4a with its physiological targets, the cyclin-dependent kinases (CDKs) CDK4 and CDK6. Assessing pathogenicity of nonsynonymous mutations is critical to evaluate melanoma risk in carriers. In the current study, we investigate 20 CDKN2A germline mutations whose effects on p16INK4A structure and function have not been previously documented (Thr18_Ala19dup, Gly23Asp, Arg24Gln, Gly35Ala, Gly35Val, Ala57Val, Ala60Val, Ala60Arg, Leu65dup, Gly67Arg, Gly67_Asn71del, Glu69Gly, Asp74Tyr, Thr77Pro, Arg80Pro, Pro81Thr, Arg87Trp, Leu97Arg, Arg99Pro, and [Leu113Leu;Pro114Ser]). By considering genetic information, the predicted impact of each variant on the protein structure, its ability to interact with CDK4 and impede cell proliferation in experimental settings, we conclude that 18 of the 20 CDKN2A variants can be classed as loss of function mutations, whereas the results for two remain ambiguous. Discriminating between mutant and neutral variants of p16INK4A not only adds to our understanding of the functionally critical residues in the protein but provides information that can be used for melanoma risk prediction. Hum Mutat 0, 1–11, 2009. © 2009 Wiley-Liss, Inc.

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