CBP and p300 histone acetyltransferases contribute to homologous recombination by transcriptionally activating the BRCA1 and RAD51 genes

PLoS One. 2012;7(12):e52810. doi: 10.1371/journal.pone.0052810. Epub 2012 Dec 20.

Abstract

Histone acetylation at DNA double-strand break (DSB) sites by CBP and p300 histone acetyltransferases (HATs) is critical for the recruitment of DSB repair proteins to chromatin. Here, we show that CBP and p300 HATs also function in DSB repair by transcriptionally activating the BRCA1 and RAD51 genes, which are involved in homologous recombination (HR), a major DSB repair system. siRNA-mediated depletion of CBP and p300 impaired HR activity and downregulated BRCA1 and RAD51 at the protein and mRNA levels. Chromatin immunoprecipitation assays showed that CBP and p300 bind to the promoter regions of the BRCA1 and RAD51 genes, and that depletion of CBP and/or p300 reduces H3 and H4 acetylation and inhibits binding of the transcription factor E2F1 to these promoters. Depletion of CBP and p300 impaired DNA damage-induced phosphorylation and chromatin binding of the single-strand DNA-binding protein RPA following BRCA1-mediated DNA end resection. Consistent with this, subsequent phosphorylation of CHK1 and activation of the G2/M damage checkpoint were also impaired. These results indicate that the HATs CBP and p300 play multiple roles in the activation of the cellular response to DSBs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylation
  • BRCA1 Protein / genetics*
  • CREB-Binding Protein / metabolism*
  • Cell Line, Tumor
  • Checkpoint Kinase 1
  • DNA Breaks, Double-Stranded
  • E2F1 Transcription Factor
  • Gene Expression Regulation, Neoplastic
  • Histones / metabolism
  • Homologous Recombination*
  • Humans
  • Promoter Regions, Genetic
  • Protein Binding
  • Protein Kinases / metabolism
  • Rad51 Recombinase / genetics*
  • Replication Protein A / metabolism
  • Signal Transduction
  • Transcriptional Activation*
  • p300-CBP Transcription Factors / metabolism*

Substances

  • BRCA1 Protein
  • E2F1 Transcription Factor
  • Histones
  • Replication Protein A
  • CREB-Binding Protein
  • p300-CBP Transcription Factors
  • p300-CBP-associated factor
  • Protein Kinases
  • CHEK1 protein, human
  • Checkpoint Kinase 1
  • Rad51 Recombinase

Grant support

This work was supported by Grants-in-Aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan for Scientific Research on Innovative Areas(22131006); from the Japan Society for the Promotion of Science for Young Scientists (B) KAKENHI (23701110); and from the National Cancer Center Research and Development Fund. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.