Dynamic in vivo interaction of DDB2 E3 ubiquitin ligase with UV-damaged DNA is independent of damage-recognition protein XPC

Martijn S. Luijsterburg, Joachim Goedhart, Jill Moser, Hanneke Kool, Bart Geverts, Adriaan B. Houtsmuller, Leon H. F. Mullenders, Wim Vermeulen, Roel van Driel*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

83 Citations (Scopus)
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Abstract

Damage DNA binding protein 2 ( DDB2) has a high affinity for UV-damaged DNA and has been implicated in the initial steps of global genome nucleotide excision repair (NER) in mammals. DDB2 binds to CUL4A and forms an E3 ubiquitin ligase. In this study, we have analyzed the properties of DDB2 and CUL4A in vivo. The majority of DDB2 and CUL4A diffuse in the nucleus with a diffusion rate consistent with a high molecular mass complex. Essentially all DDB2 binds to UV-induced DNA damage, where each molecule resides for similar to 2 minutes. After the induction of DNA damage, DDB2 is proteolytically degraded with a half-life that is two orders of magnitude larger than its residence time on a DNA lesion. This indicates that binding to damaged DNA is not the primary trigger for DDB2 breakdown. The bulk of DDB2 binds to and dissociates from DNA lesions independently of damage-recognition protein XPC. Moreover, the DDB2-containing E3 ubiquitin ligase is bound to many more damaged sites than XPC, suggesting that there is little physical interaction between the two proteins. We propose a scenario in which DDB2 prepares UV-damaged chromatin for assembly of the NER complex.
Original languageEnglish
Pages (from-to)2706-2716
Number of pages11
JournalJournal of Cell Science
Volume120
Issue number15
DOIs
Publication statusPublished - 1-Aug-2007
Externally publishedYes

Keywords

  • DNA repair
  • chromatin
  • live cell imaging
  • nuclear organization
  • nucleotide excision repair
  • NUCLEOTIDE EXCISION-REPAIR
  • PIGMENTOSUM GROUP-E
  • CYCLOBUTANE PYRIMIDINE DIMERS
  • GREEN FLUORESCENT PROTEIN
  • GLOBAL GENOMIC REPAIR
  • GROUP-E CELLS
  • BINDING-PROTEIN
  • INDUCED UBIQUITYLATION
  • GENE-PRODUCT
  • LIVING CELLS

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