Accurate completion of replication relies on the ability of cells to activate error-free recombination-mediated DNA damage bypass at sites of perturbed replication. However, as anti-recombinase activities are also recruited to replication forks, how recombination-mediated damage bypass is enabled at replication stress sites remained puzzling. Here we uncovered that the conserved SUMO-like domain-containing Saccharomyces cerevisiae protein Esc2 facilitates recombination-mediated DNA damage tolerance by allowing optimal recruitment of the Rad51 recombinase specifically at sites of perturbed replication. Mechanistically, Esc2 binds stalled replication forks and counteracts the anti-recombinase Srs2 helicase via a two-faceted mechanism involving chromatin recruitment and turnover of Srs2. Importantly, point mutations in the SUMO-like domains of Esc2 that reduce its interaction with Srs2 cause suboptimal levels of Rad51 recruitment at damaged replication forks. In conclusion, our results reveal how recombination-mediated DNA damage tolerance is locally enabled at sites of replication stress and globally prevented at undamaged replicating chromosomes.

Department of Biology Masaryk University CZ 62500 Brno Czech Republic; International Clinical Research Center Center for Biomolecular and Cellular Engineering St Anne's University Hospital Brno CZ 656 91 Brno Czech Republic

FIRC 20139 Milan Italy;

FIRC 20139 Milan Italy; National Centre for Biomolecular Research Masaryk University CZ 62500 Brno Czech Republic;

National Centre for Biomolecular Research Masaryk University CZ 62500 Brno Czech Republic; Department of Biology Masaryk University CZ 62500 Brno Czech Republic; International Clinical Research Center Center for Biomolecular and Cellular Engineering St Anne's University Hospital Brno CZ 656 91 Brno Czech Republic

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Cell Cycle. 2016;15(2):160-1. doi: 10.1080/15384101.2015.1118918. PubMed

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Esc2 promotes Mus81 complex-activity via its SUMO-like and DNA binding domains