Cell cycle regulation of DNA double-strand break end resection by Cdk1-dependent Dna2 phosphorylation

Nat Struct Mol Biol. 2011 Aug 14;18(9):1015-9. doi: 10.1038/nsmb.2105.

Abstract

DNA recombination pathways are regulated by the cell cycle to coordinate with replication. Cyclin-dependent kinase (Cdk1) promotes efficient 5' strand resection at DNA double-strand breaks (DSBs), the initial step of homologous recombination and damage checkpoint activation. The Mre11-Rad50-Xrs2 complex with Sae2 initiates resection, whereas two nucleases, Exo1 and Dna2, and the DNA helicase-topoisomerase complex Sgs1-Top3-Rmi1 generate longer ssDNA at DSBs. Using Saccharomyces cerevisiae, we provide evidence for Cdk1-dependent phosphorylation of the resection nuclease Dna2 at Thr4, Ser17 and Ser237 that stimulates its recruitment to DSBs, resection and subsequent Mec1-dependent phosphorylation. Poorly recruited dna2T4A S17A S237A and dna2ΔN248 mutant proteins promote resection only in the presence of Exo1, suggesting cross-talk between Dna2- and Exo1-dependent resection pathways.

Publication types

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

MeSH terms

  • CDC2 Protein Kinase / chemistry
  • CDC2 Protein Kinase / physiology*
  • DNA Breaks, Double-Stranded*
  • DNA Helicases / metabolism*
  • Exodeoxyribonucleases / metabolism
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Models, Genetic
  • Phosphorylation
  • Protein-Serine-Threonine Kinases / metabolism
  • Recombination, Genetic
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / metabolism
  • Saccharomyces cerevisiae Proteins / physiology*

Substances

  • Intracellular Signaling Peptides and Proteins
  • Saccharomyces cerevisiae Proteins
  • MEC1 protein, S cerevisiae
  • Protein-Serine-Threonine Kinases
  • CDC2 Protein Kinase
  • Exodeoxyribonucleases
  • exodeoxyribonuclease I
  • DNA Helicases
  • DNA2 protein, S cerevisiae