hHR23B is required for genotoxic-specific activation of p53 and apoptosis

Oncogene. 2007 Feb 22;26(8):1231-7. doi: 10.1038/sj.onc.1209865. Epub 2006 Aug 21.

Abstract

Rad23 proteins function in both DNA repair and protein stability regulation. As ubiquitinated forms of p53 are stabilized after DNA damage in concert with p53 functional activation, and human Rad23 proteins (hHR23A and B) regulate p53 stability in unstressed cells, the role of hHR23B in post-genotoxin regulation of p53 was investigated. Depletion of hHR23B by specific short interfering RNA before genotoxic exposure attenuated p53, p21 and bax induction, abrogated the accumulation of ubiquitinated p53 and suppressed apoptosis. Expression of ubiquitin derivatives with all lysines mutated except K48 or K63 demonstrated that K48-linked p53-ubiquitin conjugates were specifically induced after DNA damage. hHR23B, along with native and ubiquitinated p53, accumulated in chromatin after genotoxic exposure, and the accumulation of ubiquitinated p53 in chromatin was prevented by hHR23B depletion. Chromatin immunoprecipitation analysis demonstrated that hHR23B and p53 both localized to the p21 promoter shortly after DNA damage. hHR23B thus plays a critical role in the activation and function of p53 after specific genotoxic exposures.

Publication types

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

MeSH terms

  • Apoptosis* / genetics
  • Chromatin / chemistry
  • Chromatin / metabolism
  • Chromatin Immunoprecipitation
  • DNA / drug effects
  • DNA / radiation effects
  • DNA Damage*
  • DNA Repair Enzymes / analysis
  • DNA Repair Enzymes / genetics
  • DNA Repair Enzymes / physiology*
  • DNA-Binding Proteins / analysis
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / physiology*
  • Humans
  • Lysine / chemistry
  • Lysine / genetics
  • Mutation
  • Promoter Regions, Genetic
  • Proto-Oncogene Proteins p21(ras) / genetics
  • Proto-Oncogene Proteins p21(ras) / metabolism
  • RNA Interference
  • Tumor Suppressor Protein p53 / analysis
  • Tumor Suppressor Protein p53 / genetics
  • Tumor Suppressor Protein p53 / metabolism*
  • Ubiquitin / metabolism

Substances

  • Chromatin
  • DNA-Binding Proteins
  • Tumor Suppressor Protein p53
  • Ubiquitin
  • RAD23A protein, human
  • DNA
  • Proto-Oncogene Proteins p21(ras)
  • DNA Repair Enzymes
  • Lysine