Choreography of the DNA Damage Response: Spatiotemporal Relationships Among Checkpoint and Repair Proteins

Cell. 2004 Sep 17;118(6):699-713. doi: 10.1016/j.cell.2004.08.015.

Abstract

DNA repair is an essential process for preserving genome integrity in all organisms. In eukaryotes, recombinational repair is choreographed by multiprotein complexes that are organized into centers (foci). Here, we analyze the cellular response to DNA double-strand breaks (DSBs) and replication stress in Saccharomyces cerevisiae. The Mre11 nuclease and the ATM-related Tel1 kinase are the first proteins detected at DSBs. Next, the Rfa1 single-strand DNA binding protein relocalizes to the break and recruits other key checkpoint proteins. Later and only in S and G2 phase, the homologous recombination machinery assembles at the site. Unlike the response to DSBs, Mre11 and recombination proteins are not recruited to hydroxyurea-stalled replication forks unless the forks collapse. The cellular response to DSBs and DNA replication stress is likely directed by the Mre11 complex detecting and processing DNA ends in conjunction with Sae2 and by RP-A recognizing single-stranded DNA and recruiting additional checkpoint and repair proteins.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cell Cycle Proteins / genetics*
  • DNA / genetics
  • DNA / metabolism*
  • DNA Damage / genetics*
  • DNA Repair / genetics*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Endodeoxyribonucleases / genetics
  • Endodeoxyribonucleases / metabolism
  • Endonucleases
  • Exodeoxyribonucleases / genetics
  • Exodeoxyribonucleases / metabolism
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism
  • G2 Phase / genetics
  • Gamma Rays
  • Gene Expression Regulation, Fungal / genetics
  • Genes, cdc / physiology
  • Hydroxyurea / pharmacology
  • Intracellular Signaling Peptides and Proteins
  • Oxidative Stress / genetics
  • Protein-Serine-Threonine Kinases
  • Reaction Time / genetics
  • Recombination, Genetic / genetics
  • Replication Protein A
  • S Phase / genetics
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae / radiation effects
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism
  • Time Factors
  • Transcription Factors / genetics
  • Transcription Factors / metabolism

Substances

  • Cell Cycle Proteins
  • DNA-Binding Proteins
  • Fungal Proteins
  • Intracellular Signaling Peptides and Proteins
  • RFA1 protein, S cerevisiae
  • Replication Protein A
  • SAE2 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors
  • DNA
  • Protein-Serine-Threonine Kinases
  • TEL1 protein, S cerevisiae
  • Endodeoxyribonucleases
  • Endonucleases
  • Exodeoxyribonucleases
  • MRE11 protein, S cerevisiae
  • Hydroxyurea