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A cell‐free assay for the functional analysis of variants of the mismatch repair protein MLH1
Author(s) -
Drost Mark,
Zonneveld Jos é B.M.,
van Dijk Linda,
Morreau Hans,
Tops Carli M.,
Vasen Hans F.A.,
Wijnen Juul T.,
de Wind Niels
Publication year - 2010
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/humu.21180
Subject(s) - lynch syndrome , mlh1 , biology , dna mismatch repair , pms2 , in silico , concordance , genetics , msh2 , mutation , computational biology , gene , cancer research , bioinformatics , dna repair
Abstract The hereditary colon and endometrium cancer predisposition Lynch Syndrome (also called HNPCC) is caused by a germ‐line mutation in one of the DNA mismatch repair (MMR) genes. A significant fraction of the gene alterations detected in suspected Lynch Syndrome patients is comprised of amino acid substitutions. The relevance for cancer risk of these variants is difficult to assess, as currently no time‐ and cost‐effective, validated, and widely applicable functional assays for the measurement of MMR activity are available. Here we describe a rapid, cell‐free, and easily quantifiable MMR activity assay for the diagnostic assessment of variants of the MLH1 MMR protein. This assay allows the parallel generation and functional analysis of a series of variants of the MLH1 protein in vitro using readily available, or preprepared, reagents. Using this assay we have tested 26 MLH1 variants and of these, 15 had lost activity. These results are in concordance with those obtained from first‐generation assays and with in silico and pathology data. After its multifocal technical and clinical validation this assay could have great impact for the diagnosis and counseling of carriers of an MLH1 variant and their relatives. Hum Mutat 30:1–7, 2010. © 2010 Wiley‐Liss, Inc.