Promotion of presynaptic filament assembly by the ensemble of S. cerevisiae Rad51 paralogues with Rad52

William A. Gaines, Stephen K. Godin, Faiz F. Kabbinavar, Timsi Rao, Andrew P. VanDemark, Patrick Sung () and Kara A. Bernstein ()
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William A. Gaines: Yale University School of Medicine
Stephen K. Godin: University of Pittsburgh School of Medicine, 5117 Centre Avenue, UPCI Research Pavilion, G5.c, Pittsburgh, Pennsylvania 15217, USA
Faiz F. Kabbinavar: University of Pittsburgh School of Medicine, 5117 Centre Avenue, UPCI Research Pavilion, G5.c, Pittsburgh, Pennsylvania 15217, USA
Timsi Rao: Yale University School of Medicine
Andrew P. VanDemark: University of Pittsburgh
Patrick Sung: Yale University School of Medicine
Kara A. Bernstein: University of Pittsburgh School of Medicine, 5117 Centre Avenue, UPCI Research Pavilion, G5.c, Pittsburgh, Pennsylvania 15217, USA

Nature Communications, 2015, vol. 6, issue 1, 1-7

Abstract: Abstract The conserved budding yeast Rad51 paralogues, including Rad55, Rad57, Csm2 and Psy3 are indispensable for homologous recombination (HR)-mediated chromosome damage repair. Rad55 and Rad57 are associated in a heterodimer, while Csm2 and Psy3 form the Shu complex with Shu1 and Shu2. Here we show that Rad55 bridges an interaction between Csm2 with Rad51 and Rad52 and, using a fully reconstituted system, demonstrate that the Shu complex synergizes with Rad55–Rad57 and Rad52 to promote nucleation of Rad51 on single-stranded DNA pre-occupied by replication protein A (RPA). The csm2–F46A allele is unable to interact with Rad55, ablating the ability of the Shu complex to enhance Rad51 presynaptic filament assembly in vitro and impairing HR in vivo. Our results reveal that Rad55–Rad57, the Shu complex and Rad52 act as a functional ensemble to promote Rad51-filament assembly, which has important implications for understanding the role of the human RAD51 paralogues in Fanconi anaemia and cancer predisposition.

Date: 2015
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DOI: 10.1038/ncomms8834

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