iTRAQ-based chromatin proteomic screen reveals CHD4-dependent recruitment of MBD2 to sites of DNA damage

Biochem Biophys Res Commun. 2016 Feb 26;471(1):142-8. doi: 10.1016/j.bbrc.2016.01.162. Epub 2016 Jan 28.

Abstract

Many DNA repair proteins can be recruited to DNA damage sites upon genotoxic stress. In order to search potential DNA repair proteins involved in cellular response to mitomycin C treatment, we utilized a quantitative proteome to uncover proteins that manifest differentially enrichment in the chromatin fraction after DNA damage. 397 proteins were identified, among which many factors were shown to be involved in chromatin modification and DNA repair by GO analysis. Specifically, methyl-CpG-binding domain protein 2 (MBD2) is revealed to be recruited to DNA damage sites after laser microirradiation, which was mediated through MBD domain and MBD2 C-terminus. Additionally, the recruitment of MBD2 is dependent on poly (ADP-ribose) and chromodomain helicase DNA-binding protein 4 (CHD4). Moreover, knockdown of MBD2 by CRISPR-Cas9 technique results in MMC sensitivity in mammalian cells.

Keywords: CHD4; Chromatin fraction; DNA damage response; MBD2; PAR; iTRAQ.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autoantigens / metabolism*
  • Binding Sites
  • Chromosomal Proteins, Non-Histone / metabolism
  • DNA Damage / physiology*
  • DNA Repair / physiology*
  • DNA Repair / radiation effects
  • DNA-Binding Proteins / metabolism*
  • HeLa Cells
  • Humans
  • Mi-2 Nucleosome Remodeling and Deacetylase Complex / metabolism*
  • Peptide Mapping / methods*
  • Protein Binding
  • Proteome / metabolism*
  • Radiation Dosage
  • Tandem Mass Spectrometry / methods

Substances

  • Autoantigens
  • CHD4 protein, human
  • Chromosomal Proteins, Non-Histone
  • DNA-Binding Proteins
  • MBD2 protein, human
  • Proteome
  • Mi-2 Nucleosome Remodeling and Deacetylase Complex