Chemical proteomics reveals a γH2AX-53BP1 interaction in the DNA damage response

Nat Chem Biol. 2015 Oct;11(10):807-14. doi: 10.1038/nchembio.1908. Epub 2015 Sep 7.

Abstract

DNA double-strand break repair involves phosphorylation of histone variant H2AX ('γH2AX'), which accumulates in foci at sites of DNA damage. In current models, the recruitment of multiple DNA repair proteins to γH2AX foci depends mainly on recognition of this 'mark' by a single protein, MDC1. However, DNA repair proteins accumulate at γH2AX sites without MDC1, suggesting that other 'readers' of this mark exist. Here, we use a quantitative chemical proteomics approach to profile direct, phospho-selective γH2AX binders in native proteomes. We identify γH2AX binders, including the DNA repair mediator 53BP1, which we show recognizes γH2AX through its BRCT domains. Furthermore, we investigate the targeting of wild-type 53BP1, or a mutant form deficient in γH2AX binding, to chromosomal breaks resulting from endogenous and exogenous DNA damage. Our results show how direct recognition of γH2AX modulates protein localization at DNA damage sites, and suggest how specific chromatin mark-reader interactions contribute to essential mechanisms ensuring genome stability.

Publication types

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

MeSH terms

  • Adaptor Proteins, Signal Transducing
  • Catalytic Domain
  • Cell Cycle Proteins
  • Chromatin / metabolism
  • DNA Breaks, Double-Stranded*
  • DNA Repair*
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / metabolism
  • Genomic Instability
  • HeLa Cells
  • Histones / chemistry*
  • Histones / genetics
  • Histones / metabolism
  • Humans
  • Intracellular Signaling Peptides and Proteins / chemistry*
  • Intracellular Signaling Peptides and Proteins / genetics
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Nuclear Proteins / chemistry
  • Nuclear Proteins / metabolism
  • Point Mutation
  • Protein Binding
  • Protein Processing, Post-Translational
  • Proteomics / methods*
  • Trans-Activators / chemistry
  • Trans-Activators / metabolism
  • Tumor Suppressor p53-Binding Protein 1

Substances

  • Adaptor Proteins, Signal Transducing
  • Cell Cycle Proteins
  • Chromatin
  • DNA-Binding Proteins
  • H2AX protein, human
  • Histones
  • Intracellular Signaling Peptides and Proteins
  • MDC1 protein, human
  • Nuclear Proteins
  • TP53BP1 protein, human
  • Trans-Activators
  • Tumor Suppressor p53-Binding Protein 1

Associated data

  • PubChem-Substance/252165686
  • PubChem-Substance/252165687
  • PubChem-Substance/252165688