Nucleotide excision repair-initiating proteins bind to oxidative DNA lesions in vivo

J Cell Biol. 2012 Dec 24;199(7):1037-46. doi: 10.1083/jcb.201205149. Epub 2012 Dec 17.

Abstract

Base excision repair (BER) is the main repair pathway to eliminate abundant oxidative DNA lesions such as 8-oxo-7,8-dihydroguanine. Recent data suggest that the key transcription-coupled nucleotide excision repair factor (TC-NER) Cockayne syndrome group B (CSB) and the global genome NER-initiating factor XPC are implicated in the protection of cells against oxidative DNA damages. Our novel live-cell imaging approach revealed a strong and very rapid recruitment of XPC and CSB to sites of oxidative DNA lesions in living cells. The absence of detectable accumulation of downstream NER factors at the site of local oxidative DNA damage provide the first in vivo indication of the involvement of CSB and XPC in the repair of oxidative DNA lesions independent of the remainder of the NER reaction. Interestingly, CSB exhibited different and transcription-dependent kinetics in the two compartments studied (nucleolus and nucleoplasm), suggesting a direct transcription-dependent involvement of CSB in the repair of oxidative lesions associated with different RNA polymerases but not involving other NER proteins.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Cell Nucleolus / metabolism
  • DNA Glycosylases / metabolism
  • DNA Helicases / metabolism*
  • DNA Repair Enzymes / metabolism*
  • DNA Repair*
  • DNA-Binding Proteins / metabolism*
  • Dactinomycin / pharmacology
  • Fluorescence Recovery After Photobleaching
  • Green Fluorescent Proteins / metabolism
  • Guanine / analogs & derivatives*
  • Guanine / metabolism
  • Humans
  • Kinetics
  • Microscopy, Fluorescence
  • Nucleic Acid Synthesis Inhibitors / pharmacology
  • Oxidation-Reduction
  • Poly-ADP-Ribose Binding Proteins
  • Protein Binding
  • Protein Transport
  • Recombinant Fusion Proteins / metabolism
  • Transcription, Genetic / drug effects

Substances

  • DNA-Binding Proteins
  • Nucleic Acid Synthesis Inhibitors
  • Poly-ADP-Ribose Binding Proteins
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • XPC protein, human
  • Dactinomycin
  • 8-hydroxyguanine
  • Guanine
  • DNA Glycosylases
  • oxoguanine glycosylase 1, human
  • DNA Helicases
  • ERCC6 protein, human
  • DNA Repair Enzymes