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; Kara A Bernstein

doi:10.1038/ncomms8834

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

Nat Commun. 2015 Jul 28:6:7834. doi: 10.1038/ncomms8834.

Authors

William A Gaines¹, Stephen K Godin², Faiz F Kabbinavar², Timsi Rao¹, Andrew P VanDemark³, Patrick Sung¹, Kara A Bernstein²

Affiliations

¹ Department of Molecular Biochemistry and Biophysics, Yale University School of Medicine, New Haven, Conneticut 06510, USA.
² Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, 5117 Centre Avenue, UPCI Research Pavilion, G5.c, Pittsburgh, Pennsylvania 15217, USA.
³ Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, USA.

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.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Adenosine Triphosphatases / metabolism*
Cell Cycle Proteins / metabolism
DNA Repair Enzymes / metabolism*
DNA, Single-Stranded / metabolism*
DNA-Binding Proteins / metabolism*
In Vitro Techniques
Microscopy, Electron
Nuclear Proteins / metabolism
Rad51 Recombinase / metabolism*
Rad52 DNA Repair and Recombination Protein / metabolism*
Recombinational DNA Repair*
Replication Protein A / metabolism*
Saccharomyces cerevisiae
Saccharomyces cerevisiae Proteins / metabolism*
Schizosaccharomyces pombe Proteins / metabolism

Substances

Cell Cycle Proteins
Csm2 protein, S cerevisiae
DNA, Single-Stranded
DNA-Binding Proteins
Nuclear Proteins
PSY3 protein, S cerevisiae
RAD52 protein, S cerevisiae
RAD55 protein, S cerevisiae
Rad52 DNA Repair and Recombination Protein
Replication Protein A
Saccharomyces cerevisiae Proteins
Schizosaccharomyces pombe Proteins
Shu1 protein, S cerevisiae
shu2 protein, S cerevisiae
RAD51 protein, S cerevisiae
Rad51 Recombinase
Adenosine Triphosphatases
RAD57 protein, S cerevisiae
DNA Repair Enzymes

Abstract

Publication types

MeSH terms

Substances

Grants and funding