The RecQ DNA helicase Rqh1 constrains Exonuclease 1-dependent recombination at stalled replication forks

Sci Rep. 2016 Mar 9:6:22837. doi: 10.1038/srep22837.

Abstract

DNA double-strand break (DSB) repair by homologous recombination (HR) involves resection of the break to expose a 3' single-stranded DNA tail. In budding yeast, resection occurs in two steps: initial short-range resection, performed by Mre11-Rad50-Xrs2 and Sae2; and long-range resection catalysed by either Exo1 or Sgs1-Dna2. Here we use genetic assays to investigate the importance of Exo1 and the Sgs1 homologue Rqh1 for DNA repair and promotion of direct repeat recombination in the fission yeast Schizosaccharomyces pombe. We find that Exo1 and Rqh1 function in alternative redundant pathways for promoting survival following replication fork breakage. Exo1 promotes replication fork barrier-induced direct repeat recombination but intriguingly limits recombination induced by fork breakage. Direct repeat recombination induced by ultraviolet light depends on either Exo1 or Rqh1. Finally, we show that Rqh1 plays a major role in limiting Exo1-dependent direct repeat recombination induced by replication fork stalling but only a minor role in constraining recombination induced by fork breakage. The implications of our findings are discussed in the context of the benefits that long-range resection may bring to processing perturbed replication forks.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Helicases / metabolism*
  • DNA Repair
  • DNA Replication*
  • Exodeoxyribonucleases / antagonists & inhibitors*
  • Recombination, Genetic*
  • Schizosaccharomyces / enzymology*
  • Schizosaccharomyces / genetics*
  • Schizosaccharomyces pombe Proteins / metabolism*

Substances

  • Schizosaccharomyces pombe Proteins
  • Exodeoxyribonucleases
  • exodeoxyribonuclease I
  • DNA Helicases
  • Rqh1 protein, S pombe