Histone Deposition Promotes Recombination-Dependent Replication at Arrested Forks

PLoS Genet. 2019 Oct 4;15(10):e1008441. doi: 10.1371/journal.pgen.1008441. eCollection 2019 Oct.

Abstract

Replication stress poses a serious threat to genome stability. Recombination-Dependent-Replication (RDR) promotes DNA synthesis resumption from arrested forks. Despite the identification of chromatin restoration pathways after DNA repair, crosstalk coupling RDR and chromatin assembly is largely unexplored. The fission yeast Chromatin Assembly Factor-1, CAF-1, is known to promote RDR. Here, we addressed the contribution of histone deposition to RDR. We expressed a mutated histone, H3-H113D, to genetically alter replication-dependent chromatin assembly by destabilizing (H3-H4)2 tetramer. We established that DNA synthesis-dependent histone deposition, by CAF-1 and Asf1, promotes RDR by preventing Rqh1-mediated disassembly of joint-molecules. The recombination factor Rad52 promotes CAF-1 binding to sites of recombination-dependent DNA synthesis, indicating that histone deposition occurs downstream Rad52. Histone deposition and Rqh1 activity act synergistically to promote cell resistance to camptothecin, a topoisomerase I inhibitor that induces replication stress. Moreover, histone deposition favors non conservative recombination events occurring spontaneously in the absence of Rqh1, indicating that the stabilization of joint-molecules by histone deposition also occurs independently of Rqh1 activity. These results indicate that histone deposition plays an active role in promoting RDR, a benefit counterbalanced by stabilizing at-risk joint-molecules for genome stability.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle Proteins / metabolism
  • Chromatin Assembly and Disassembly*
  • DNA Helicases / metabolism
  • DNA Replication*
  • Genomic Instability*
  • Histones / genetics
  • Histones / metabolism*
  • Molecular Chaperones / metabolism
  • Mutation
  • Protein Multimerization / genetics
  • Rad52 DNA Repair and Recombination Protein / metabolism
  • Recombination, Genetic*
  • Ribonucleases / metabolism
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae Proteins / metabolism
  • Schizosaccharomyces / genetics
  • Schizosaccharomyces pombe Proteins / metabolism

Substances

  • ASF1 protein, S cerevisiae
  • Cell Cycle Proteins
  • Cia1 protein, S pombe
  • Histones
  • Molecular Chaperones
  • Rad52 DNA Repair and Recombination Protein
  • Saccharomyces cerevisiae Proteins
  • Schizosaccharomyces pombe Proteins
  • Pop2 protein, S pombe
  • Ribonucleases
  • POP2 protein, S cerevisiae
  • DNA Helicases
  • Rqh1 protein, S pombe

Grant support

This study was supported by grants from the Institut Curie, the CNRS, the Fondation ARC pour la recherche sur le cancer (https://www.fondation-arc.org/), the Fondation Ligue (comité Essonne, https://www.ligue-cancer.net/), l’Agence Nationale de la Recherche [grant number ANR-14-CE10-0010-01, https://anr.fr/en/], the Institut National du Cancer [grant number 2016-1-PLBIO-03-ICR-1, https://en.e-cancer.fr/] and the Fondation pour la Recherche Médicale [grant number Equipe FRM DEQ20160334889, https://www.frm.org/en]. ATS and AAS were funded by the Institut Curie international PhD program, and a French governmental fellowship, respectively. ATS was supported by a 4th year PhD fellowship from Fondation pour la Recherche Médicale [grant number FDT20160435131]. The funders had no role in study design, data collection and analysis, the decision to publish, or preparation of the manuscript.