Break-induced replication promotes formation of lethal joint molecules dissolved by Srs2

Nat Commun. 2017 Nov 27;8(1):1790. doi: 10.1038/s41467-017-01987-2.

Abstract

Break-induced replication (BIR) is a DNA double-strand break repair pathway that leads to genomic instabilities similar to those observed in cancer. BIR proceeds by a migrating bubble where asynchrony between leading and lagging strand synthesis leads to accumulation of long single-stranded DNA (ssDNA). It remains unknown how this ssDNA is prevented from unscheduled pairing with the template, which can lead to genomic instability. Here, we propose that uncontrolled Rad51 binding to this ssDNA promotes formation of toxic joint molecules that are counteracted by Srs2. First, Srs2 dislodges Rad51 from ssDNA preventing promiscuous strand invasions. Second, it dismantles toxic intermediates that have already formed. Rare survivors in the absence of Srs2 rely on structure-specific endonucleases, Mus81 and Yen1, that resolve toxic joint-molecules. Overall, we uncover a new feature of BIR and propose that tight control of ssDNA accumulated during this process is essential to prevent its channeling into toxic structures threatening cell viability.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Cell Survival / genetics
  • DNA Breaks, Double-Stranded
  • DNA Helicases / physiology*
  • DNA Repair / genetics*
  • DNA Replication / physiology*
  • DNA, Single-Stranded / genetics
  • DNA, Single-Stranded / metabolism*
  • DNA-Binding Proteins / metabolism
  • Endonucleases / metabolism
  • Holliday Junction Resolvases / metabolism
  • Protein Binding / physiology
  • Rad51 Recombinase / physiology
  • Saccharomyces cerevisiae / physiology*
  • Saccharomyces cerevisiae Proteins / metabolism
  • Saccharomyces cerevisiae Proteins / physiology*

Substances

  • DNA, Single-Stranded
  • DNA-Binding Proteins
  • Saccharomyces cerevisiae Proteins
  • SRS2 protein, S cerevisiae
  • RAD51 protein, S cerevisiae
  • Rad51 Recombinase
  • Endonucleases
  • MUS81 protein, S cerevisiae
  • Holliday Junction Resolvases
  • Yen1 protein, S cerevisiae
  • DNA Helicases