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Title: Rad52 Restrains Resection at DNA Double-Strand Break Ends in Yeast.

Authors: Yan, Zhenxin; Xue, Chaoyou; Kumar, Sandeep; Crickard, J Brooks; Yu, Yang; Wang, Weibin; Pham, Nhung; Li, Yuxi; Niu, Hengyao; Sung, Patrick; Greene, Eric C; Ira, Grzegorz

Published In Mol Cell, (2019 12 05)

Abstract: Rad52 is a key factor for homologous recombination (HR) in yeast. Rad52 helps assemble Rad51-ssDNA nucleoprotein filaments that catalyze DNA strand exchange, and it mediates single-strand DNA annealing. We find that Rad52 has an even earlier function in HR in restricting DNA double-stranded break ends resection that generates 3' single-stranded DNA (ssDNA) tails. In fission yeast, Exo1 is the primary resection nuclease, with the helicase Rqh1 playing a minor role. We demonstrate that the choice of two extensive resection pathways is regulated by Rad52. In rad52 cells, the resection rate increases from ∼3-5 kb/h up to ∼10-20 kb/h in an Rqh1-dependent manner, while Exo1 becomes dispensable. Budding yeast Rad52 similarly inhibits Sgs1-dependent resection. Single-molecule analysis with purified budding yeast proteins shows that Rad52 competes with Sgs1 for DNA end binding and inhibits Sgs1 translocation along DNA. These results identify a role for Rad52 in limiting ssDNA generated by end resection.

PubMed ID: 31542296 Exiting the NIEHS site

MeSH Terms: DNA Breaks, Double-Stranded*; DNA Breaks, Single-Stranded*; DNA Helicases/genetics; DNA Helicases/metabolism; DNA Repair*; DNA, Fungal/genetics; DNA, Fungal/metabolism*; DNA-Binding Proteins/genetics; DNA-Binding Proteins/metabolism*; Exodeoxyribonucleases/genetics; Exodeoxyribonucleases/metabolism; Gene Expression Regulation, Fungal; Kinetics; Mutation; Protein Domains; Protein Transport; Rad52 DNA Repair and Recombination Protein/genetics; Rad52 DNA Repair and Recombination Protein/metabolism*; RecQ Helicases/genetics; RecQ Helicases/metabolism; Saccharomyces cerevisiae Proteins/genetics; Saccharomyces cerevisiae Proteins/metabolism*; Saccharomyces cerevisiae/enzymology*; Saccharomyces cerevisiae/genetics; Schizosaccharomyces pombe Proteins/genetics; Schizosaccharomyces pombe Proteins/metabolism*; Schizosaccharomyces/enzymology*; Schizosaccharomyces/genetics

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