Germline missense mutations affecting a single BRCA2 allele predispose humans to cancer. Here we identify a protein-targeting mechanism that is disrupted by the cancer-associated mutation, BRCA2D2723H, and that controls the nuclear localization of BRCA2 and its cargo, the recombination enzyme RAD51. A nuclear export signal (NES) in BRCA2 is masked by its interaction with a partner protein, DSS1, such that point mutations impairing BRCA2-DSS1 binding render BRCA2 cytoplasmic. In turn, cytoplasmic mislocalization of mutant BRCA2 inhibits the nuclear retention of RAD51 by exposing a similar NES in RAD51 that is usually obscured by the BRCA2-RAD51 interaction. Thus, a series of NES-masking interactions localizes BRCA2 and RAD51 in the nucleus. Notably, BRCA2D2723H decreases RAD51 nuclear retention even when wild-type BRCA2 is also present. Our findings suggest a mechanism for the regulation of the nucleocytoplasmic distribution of BRCA2 and RAD51 and its impairment by a heterozygous disease-associated mutation.
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We acknowledge the generous gifts of WT and BRCA2D2723H ES cells from S. Sharan (US National Institutes of Health) and sYFP cDNA from T. Gadella (University of Amsterdam). We thank D. Görlich (Max Planck Institute) for generously providing constructs encoding RanQ69L and CRM1 and for information concerning assay conditions. We thank members of the Venkitaraman laboratory for technical assistance and constructive discussions and M. Goode for help with the feeder-free culture for ES cell cultures. A.D.J. was supported by a career development fellowship from the UK MRC Cancer Unit. Work in the laboratory of C.F.K. was funded by the UK Biosciences and Biotechnology Research Council, and work in the laboratory of A.R.V. was funded by the UK MRC.
The authors declare no competing financial interests.
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Jeyasekharan, A., Liu, Y., Hattori, H. et al. A cancer-associated BRCA2 mutation reveals masked nuclear export signals controlling localization. Nat Struct Mol Biol 20, 1191–1198 (2013). https://doi.org/10.1038/nsmb.2666
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