Ku prevents Exo1 and Sgs1-dependent resection of DNA ends in the absence of a functional MRX complex or Sae2

EMBO J. 2010 Oct 6;29(19):3358-69. doi: 10.1038/emboj.2010.193. Epub 2010 Aug 20.

Abstract

In this study, we investigate the interplay between Ku, a central non-homologous end-joining component, and the Mre11-Rad50-Xrs2 (MRX) complex and Sae2, end-processing factors crucial for initiating 5'-3' resection of double-strand break (DSB) ends. We show that in the absence of end protection by Ku, the requirement for the MRX complex is bypassed and resection is executed by Exo1. In contrast, both the Exo1 and Sgs1 resection pathways contribute to DSB processing in the absence of Ku and Sae2 or when the MRX complex is intact, but functionally compromised by elimination of the Mre11 nuclease activity. The ionizing radiation sensitivity of a mutant defective for extensive resection (exo1Δ sgs1Δ) cannot be suppressed by the yku70Δ mutation, indicating that Ku suppression is specific to the initiation of resection. We provide evidence that replication-associated DSBs need to be processed by Sae2 for repair by homologous recombination unless Ku is absent. Finally, we show that the presence of Ku exacerbates DNA end-processing defects established in the sae2Δ sgs1Δ mutant, leading to its lethality.

Publication types

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

MeSH terms

  • Blotting, Southern
  • DNA Breaks, Double-Stranded*
  • DNA Repair*
  • DNA-Binding Proteins / metabolism*
  • Endodeoxyribonucleases / metabolism
  • Endonucleases / metabolism
  • Exodeoxyribonucleases / metabolism*
  • Gamma Rays
  • Multiprotein Complexes / metabolism*
  • RecQ Helicases / metabolism*
  • Saccharomyces cerevisiae
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Telomere / genetics

Substances

  • DNA-Binding Proteins
  • Multiprotein Complexes
  • RAD50 protein, S cerevisiae
  • SAE2 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • XRS2 protein, S cerevisiae
  • high affinity DNA-binding factor, S cerevisiae
  • Endodeoxyribonucleases
  • Endonucleases
  • Exodeoxyribonucleases
  • MRE11 protein, S cerevisiae
  • exodeoxyribonuclease I
  • SGS1 protein, S cerevisiae
  • RecQ Helicases