Homologous recombination (HR) protects replication forks (RFs) and repairs DNA double-strand breaks (DSBs). Within HR, BRCA2 regulates RAD51 via two interaction regions: the BRC repeats to form filaments on single-stranded DNA and exon 27 (Ex27) to stabilize the filament. Here, we identified a RAD51 S181P mutant that selectively disrupted the RAD51-Ex27 association while maintaining interaction with BRC repeat and proficiently forming filaments capable of DNA binding and strand invasion. Interestingly, RAD51 S181P was defective for RF protection/restart but proficient for DSB repair. Our data suggest that Ex27-mediated stabilization of RAD51 filaments is required for the protection of RFs, while it seems dispensable for the repair of DSBs.

Center for Genomic Integrity Institute for Basic Science Ulsan 44919 Republic of Korea

Department of Biology Masaryk University 625 00 Brno Czech Republic

Department of Genetics Albert Einstein College of Medicine of Yeshiva University Bronx NY 10461 USA

Department of Molecular Medicine The Barshop Institute for Longevity and Aging Studies The Cancer Therapy Research Center UT Health San Antonio San Antonio TX 78229 USA

National Centre for Biomolecular Research Masaryk University 625 00 Brno Czech Republic

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