Rad18 is a transcriptional target of E2F3

Cell Cycle. 2012 Mar 15;11(6):1131-41. doi: 10.4161/cc.11.6.19558. Epub 2012 Mar 15.

Abstract

The E2F family of transcription factors responds to a variety of intracellular and extracellular signals and, as such, are key regulators of cell growth, differentiation and cell death. The cellular response to DNA damage is a multistep process generally involving the initial detection of DNA damage, propagation of signals via posttranslational modifications (e.g., phosphorylation and ubiquitination) and, finally, the implementation of a response. We have previously reported that E2F3 can be induced by DNA damage, and that it plays an important role in DNA damage-induced apoptosis. Here, we demonstrate that E2F3 knockdown compromises two canonical DNA damage modification events, the ubiquitination of H2AX and PCNA. We find that the defect in these posttranscriptional modifications after E2F3 knockdown is due to reduced expression of important DNA damage responsive ubiquitin ligases. We characterized the regulation of one of these ligases, Rad18, and we demonstrated that E2F3 associates with the Rad18 promoter and directly controls its activity. Furthermore, we find that ectopic expression of Rad18 is sufficient to rescue the PCNA ubiquitination defect resulting from E2F3 knockdown. Our study reveals a novel facet of E2F3's control of the DNA damage response.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis
  • Cell Line, Tumor
  • Cisplatin / pharmacology
  • DNA Damage*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • E2F1 Transcription Factor / genetics
  • E2F1 Transcription Factor / metabolism
  • E2F3 Transcription Factor / genetics
  • E2F3 Transcription Factor / metabolism*
  • G1 Phase Cell Cycle Checkpoints
  • Gene Expression Profiling
  • Gene Expression Regulation, Neoplastic*
  • Gene Knockdown Techniques
  • Histones / genetics
  • Histones / metabolism
  • Humans
  • Phosphorylation
  • Proliferating Cell Nuclear Antigen / genetics
  • Proliferating Cell Nuclear Antigen / metabolism
  • Promoter Regions, Genetic
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Transfection
  • Ubiquitin-Protein Ligases
  • Ubiquitination

Substances

  • DNA-Binding Proteins
  • E2F1 Transcription Factor
  • E2F1 protein, human
  • E2F3 Transcription Factor
  • E2F3 protein, human
  • H2AX protein, human
  • Histones
  • Proliferating Cell Nuclear Antigen
  • RAD18 protein, human
  • RNA, Small Interfering
  • Ubiquitin-Protein Ligases
  • Cisplatin