Break-induced ATR and Ddb1-Cul4(Cdt)² ubiquitin ligase-dependent nucleotide synthesis promotes homologous recombination repair in fission yeast

Genes Dev. 2010 Dec 1;24(23):2705-16. doi: 10.1101/gad.1970810.

Abstract

Nucleotide synthesis is a universal response to DNA damage, but how this response facilitates DNA repair and cell survival is unclear. Here we establish a role for DNA damage-induced nucleotide synthesis in homologous recombination (HR) repair in fission yeast. Using a genetic screen, we found the Ddb1-Cul4(Cdt)² ubiquitin ligase complex and ribonucleotide reductase (RNR) to be required for HR repair of a DNA double-strand break (DSB). The Ddb1-Cul4(Cdt)² ubiquitin ligase complex is required for degradation of Spd1, an inhibitor of RNR in fission yeast. Accordingly, deleting spd1(+) suppressed the DNA damage sensitivity and the reduced HR efficiency associated with loss of ddb1(+) or cdt2(+). Furthermore, we demonstrate a role for nucleotide synthesis in postsynaptic gap filling of resected ssDNA ends during HR repair. Finally, we define a role for Rad3 (ATR) in nucleotide synthesis and HR through increasing Cdt2 nuclear levels in response to DNA damage. Our findings support a model in which break-induced Rad3 and Ddb1-Cul4(Cdt)² ubiquitin ligase-dependent Spd1 degradation and RNR activation promotes postsynaptic ssDNA gap filling during HR repair.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / metabolism*
  • Cell Cycle Proteins / metabolism*
  • Checkpoint Kinase 2
  • DNA Breaks, Double-Stranded
  • DNA Repair
  • DNA-Binding Proteins / metabolism*
  • Gene Deletion
  • Nucleotides / metabolism
  • Protein Kinases / metabolism*
  • Recombination, Genetic
  • Ribonucleotide Reductases / metabolism
  • Schizosaccharomyces / genetics*
  • Schizosaccharomyces / metabolism*
  • Schizosaccharomyces pombe Proteins / metabolism*

Substances

  • Adaptor Proteins, Signal Transducing
  • Cdt2 protein, S pombe
  • Cell Cycle Proteins
  • DNA-Binding Proteins
  • Ddb1 protein, S pombe
  • Nucleotides
  • Schizosaccharomyces pombe Proteins
  • Spd1 protein, S pombe
  • Ribonucleotide Reductases
  • Protein Kinases
  • Checkpoint Kinase 2
  • rad3 protein, S pombe