@article{a1ad7c89ecc045a6a2871de0e2be02cb,
title = "Functional analysis of genetic variants in the high-risk breast cancer susceptibility gene PALB2",
abstract = "Heterozygous carriers of germ-line loss-of-function variants in the DNA repair gene PALB2 are at a highly increased lifetime risk for developing breast cancer. While truncating variants in PALB2 are known to increase cancer risk, the interpretation of missense variants of uncertain significance (VUS) is in its infancy. Here we describe the development of a relatively fast and easy cDNA-based system for the semi high-throughput functional analysis of 48 VUS in human PALB2. By assessing the ability of PALB2 VUS to rescue the DNA repair and checkpoint defects in Palb2 knockout mouse embryonic stem (mES) cells, we identify various VUS in PALB2 that impair its function. Three VUS in the coiled-coil domain of PALB2 abrogate the interaction with BRCA1, whereas several VUS in the WD40 domain dramatically reduce protein stability. Thus, our functional assays identify damaging VUS in PALB2 that may increase cancer risk.",
author = "Boonen, {Rick A.C.M.} and Am{\'e}lie Rodrigue and Chantal Stoepker and Wiegant, {Wouter W.} and Bas Vroling and Milan Sharma and Rother, {Magdalena B.} and Nandi Celosse and Vreeswijk, {Maaike P.G.} and Fergus Couch and Jacques Simard and Peter Devilee and Masson, {Jean Yves} and {van Attikum}, Haico",
note = "Funding Information: The authors would like to thank Jos Jonkers and Peter Bouwman for providing the pTT5-Puro (RMCE acceptor cassette), pRNA-251-MCS-RMCE (RMCE exchange vector) and pCMV-Red-I-SceI constructs, as well as Dan Durocher and Bing Xia for sharing BRCA1 and PALB2 antibodies, respectively. We are grateful to Maria Jasin and Francis Stewart for sharing the DR-GFP reporter and FlpO constructs, respectively. We thank Patrick van Vliet and Richard Lemmers for help with the Southern blot analysis, and Cell Signaling for providing antibodies directed against PALB2 prior to commercialization. Finally, we would like to thank Jamie Allen and the BRIDGES consortium for providing PALB2 variants. This work was financially supported by the Government of Canada through Genome Canada and the Canadian Institutes of Health Research, the Minist{\`e}re de l{\textquoteright}{\'E}conomie, de la Science et de l{\textquoteright}Innovation du Qu{\'e}bec through Genome Qu{\'e}bec and the Quebec Breast Cancer Foundation (J.S. and J.-Y.M.), as well as by grants from the Minist{\`e}re de l'{\'E}conomie, de la Science et de l{\textquoteright}Innovation du Qu{\'e}bec through the PSR-SIIRI-949 program (J.S. and J.-Y.M), the CIHR (Foundation grant to J.-Y.M), European Union (BRIDGES grant to P.D., M.V., and H.v.A.) and the Dutch Cancer Society (P.D. and H.v.A.). Publisher Copyright: {\textcopyright} 2019, The Author(s).",
year = "2019",
month = dec,
day = "1",
doi = "10.1038/s41467-019-13194-2",
language = "English (US)",
volume = "10",
journal = "Nature Communications",
issn = "2041-1723",
publisher = "Nature Publishing Group",
number = "1",
}