Ku stabilizes replication forks in the absence of Brc1

PLoS One. 2015 May 12;10(5):e0126598. doi: 10.1371/journal.pone.0126598. eCollection 2015.

Abstract

DNA replication errors are a major source of genome instability in all organisms. In the fission yeast Schizosaccharomyces pombe, the DNA damage response protein Brc1 binds phospho-histone H2A (γH2A)-marked chromatin during S-phase, but how Brc1 protects genome integrity remains unclear. Here we report that the non-homologous end-joining (NHEJ) protein Ku becomes critical for survival of replication stress in brc1∆ cells. Ku's protective activity in brc1∆ cells does not involve its canonical NHEJ function or its roles in protecting telomeres or shielding DNA ends from Exo1 exonuclease. In brc1∆ pku80∆ cells, nuclear foci of Rad52 homologous recombination (HR) protein increase and Mus81-Eme1 Holliday junction resolvase becomes critical, indicating increased replication fork instability. Ku's localization at a ribosomal DNA replication fork barrier associated with frequent replisome-transcriptosome collisions increases in brc1∆ cells and increased collisions correlate with an enhanced requirement for Brc1. These data indicate that Ku stabilizes replication forks in the absence of Brc1.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Antigens, Nuclear / metabolism*
  • DNA Damage
  • DNA End-Joining Repair
  • DNA Replication
  • DNA, Fungal / metabolism*
  • DNA-Binding Proteins / metabolism*
  • Genomic Instability
  • Ku Autoantigen
  • S Phase
  • Schizosaccharomyces / genetics*
  • Schizosaccharomyces pombe Proteins / metabolism*

Substances

  • Antigens, Nuclear
  • Brc1 protein, S pombe
  • DNA, Fungal
  • DNA-Binding Proteins
  • Schizosaccharomyces pombe Proteins
  • Ku Autoantigen