title: Local regulation of the Srs2 helicase by the SUMO-like domain protein Esc2 promotes recombination at sites of stalled replication.
abstract: 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.
authors.affiliation: FIRC (Fondazione Italiana per la Ricerca sul Cancro) Institute of Molecular Oncology (IFOM), 20139 Milan, Italy;
authors.forename: Madhusoodanan
authors.initials: M
authors.lastname: Urulangodi
authors.affiliation: FIRC (Fondazione Italiana per la Ricerca sul Cancro) Institute of Molecular Oncology (IFOM), 20139 Milan, Italy; National Centre for Biomolecular Research, Masaryk University, CZ-62500 Brno, Czech Republic;
authors.forename: Marek
authors.initials: M
authors.lastname: Sebesta
authors.affiliation: FIRC (Fondazione Italiana per la Ricerca sul Cancro) Institute of Molecular Oncology (IFOM), 20139 Milan, Italy;
authors.forename: Demis
authors.initials: D
authors.lastname: Menolfi
authors.affiliation: FIRC (Fondazione Italiana per la Ricerca sul Cancro) Institute of Molecular Oncology (IFOM), 20139 Milan, Italy;
authors.forename: Barnabas
authors.initials: B
authors.lastname: Szakal
authors.affiliation: FIRC (Fondazione Italiana per la Ricerca sul Cancro) Institute of Molecular Oncology (IFOM), 20139 Milan, Italy;
authors.forename: Julie
authors.initials: J
authors.lastname: Sollier
authors.affiliation: 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.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Sisakova
authors.affiliation: 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.
authors.forename: Lumir
authors.initials: L
authors.lastname: Krejci
authors.affiliation: FIRC (Fondazione Italiana per la Ricerca sul Cancro) Institute of Molecular Oncology (IFOM), 20139 Milan, Italy;
authors.forename: Dana
authors.initials: D
authors.lastname: Branzei
mesh_terms: D018797:Cell Cycle Proteins; D002843:Chromatin / Q000378:metabolism; D004249:DNA Damage / Q000235:genetics; D004265:DNA Helicases / Q000235:genetics* / Q000378:metabolism; D004261:DNA Replication / Q000235:genetics*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D017354:Point Mutation; D011485:Protein Binding; D051135:Rad51 Recombinase / Q000378:metabolism; D011995:Recombination, Genetic / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism*
chemical_list: D018797:Cell Cycle Proteins; D002843:Chromatin; C532918:Esc2 protein, S cerevisiae; D009687:Nuclear Proteins; D029701:Saccharomyces cerevisiae Proteins; C061376:SRS2 protein, S cerevisiae; C495151:RAD51 protein, S cerevisiae; D051135:Rad51 Recombinase; D004265:DNA Helicases
keywords: DNA damage tolerance; SUMO; genotoxic stress; recombination; replication