Sgs1 helicase and two nucleases Dna2 and Exo1 resect DNA double-strand break ends

Cell. 2008 Sep 19;134(6):981-94. doi: 10.1016/j.cell.2008.08.037.

Abstract

Formation of single-strand DNA (ssDNA) tails at a double-strand break (DSB) is a key step in homologous recombination and DNA-damage signaling. The enzyme(s) producing ssDNA at DSBs in eukaryotes remain unknown. We monitored 5'-strand resection at inducible DSB ends in yeast and identified proteins required for two stages of resection: initiation and long-range 5'-strand resection. We show that the Mre11-Rad50-Xrs2 complex (MRX) initiates 5' degradation, whereas Sgs1 and Dna2 degrade 5' strands exposing long 3' strands. Deletion of SGS1 or DNA2 reduces resection and DSB repair by single-strand annealing between distant repeats while the remaining long-range resection activity depends on the exonuclease Exo1. In exo1Deltasgs1Delta double mutants, the MRX complex together with Sae2 nuclease generate, in a stepwise manner, only few hundred nucleotides of ssDNA at the break, resulting in inefficient gene conversion and G2/M damage checkpoint arrest. These results provide important insights into the early steps of DSB repair in eukaryotes.

Publication types

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

MeSH terms

  • Adenosine Triphosphatases / metabolism
  • Animals
  • DNA Breaks, Double-Stranded*
  • DNA Helicases / genetics
  • DNA Helicases / metabolism*
  • DNA Repair*
  • Endodeoxyribonucleases / metabolism
  • Exodeoxyribonucleases / metabolism*
  • Gene Conversion
  • Protein Structure, Tertiary
  • RecQ Helicases / genetics
  • RecQ Helicases / metabolism*
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*

Substances

  • Saccharomyces cerevisiae Proteins
  • XRS2 protein, S cerevisiae
  • Endodeoxyribonucleases
  • Exodeoxyribonucleases
  • MRE11 protein, S cerevisiae
  • exodeoxyribonuclease I
  • Adenosine Triphosphatases
  • RAD5 protein, S cerevisiae
  • SGS1 protein, S cerevisiae
  • DNA Helicases
  • DNA2 protein, S cerevisiae
  • RecQ Helicases