Versatile DNA damage detection by the global genome nucleotide excision repair protein XPC

J Cell Sci. 2008 Sep 1;121(Pt 17):2850-9. doi: 10.1242/jcs.031708. Epub 2008 Aug 5.

Abstract

To investigate how the nucleotide excision repair initiator XPC locates DNA damage in mammalian cell nuclei we analyzed the dynamics of GFP-tagged XPC. Photobleaching experiments showed that XPC constantly associates with and dissociates from chromatin in the absence of DNA damage. DNA-damaging agents retard the mobility of XPC, and UV damage has the most pronounced effect on the mobility of XPC-GFP. XPC exhibited a surprising distinct dynamic behavior and subnuclear distribution compared with other NER factors. Moreover, we uncovered a novel regulatory mechanism for XPC. Under unchallenged conditions, XPC is continuously exported from and imported into the nucleus, which is impeded when NER lesions are present. XPC is omnipresent in the nucleus, allowing a quick response to genotoxic stress. To avoid excessive DNA probing by the low specificity of the protein, the steady-state level in the nucleus is controlled by nucleus-cytoplasm shuttling, allowing temporally higher concentrations of XPC in the nucleus under genotoxic stress conditions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Cell Line
  • Cell Nucleus / metabolism
  • Cell Nucleus / radiation effects
  • Cell Survival / radiation effects
  • DNA Damage*
  • DNA Repair* / radiation effects
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / metabolism*
  • Fibroblasts / metabolism
  • Fibroblasts / radiation effects
  • Fluorescence Recovery After Photobleaching
  • Genome / genetics*
  • Genome / radiation effects
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Kinetics
  • Models, Biological
  • Molecular Sequence Data
  • Mutant Proteins / metabolism
  • Protein Binding / radiation effects
  • Protein Transport / radiation effects
  • Recombinant Fusion Proteins / metabolism
  • Transcription Factor TFIIH / metabolism
  • Ultraviolet Rays
  • Xeroderma Pigmentosum Group A Protein / metabolism

Substances

  • DNA-Binding Proteins
  • Mutant Proteins
  • Recombinant Fusion Proteins
  • Xeroderma Pigmentosum Group A Protein
  • Green Fluorescent Proteins
  • Transcription Factor TFIIH
  • XPC protein, human