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Visualization of DNA Repair Proteins Interaction by Immunofluorescence
Published on: June 26, 2020
RAD51 separation of function mutation disables replication fork maintenance but preserves DSB repair
Mi Young Son1, Ondrej Belan2, Mario Spirek2,3
1Department 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.
None:
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.
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