UV-DDB-dependent regulation of nucleotide excision repair kinetics in living cells

Ryotaro Nishi, Sergey Alekseev, Christoffel Dinant, Deborah Hoogstraten, Adriaan B. Houtsmuller, Jan H.J. Hoeijmakers, Wim Vermeulen, Fumio Hanaoka, Kaoru Sugasawa

Research output: Contribution to journalArticlepeer-review

65 Citations (Scopus)

Abstract

Although the basic principle of nucleotide excision repair (NER), which can eliminate various DNA lesions, have been dissected at the genetic, biochemical and cellular levels, the important in vivo regulation of the critical damage recognition step is poorly understood. Here we analyze the in vivo dynamics of the essential NER damage recognition factor XPC fused to the green fluorescence protein (GFP). Fluorescence recovery after photobleaching analysis revealed that the UV-induced transient immobilization of XPC, reflecting its actual engagement in NER, is regulated in a biphasic manner depending on the number of (6-4) photoproducts and titrated by the number of functional UV-DDB molecules. A similar biphasic UV-induced immobilization of TFIIH was observed using XPB-GFP. Surprisingly, subsequent integration of XPA into the NER complex appears to follow only the low UV dose immobilization of XPC. Our results indicate that when only a small number of (6-4) photoproducts are generated, the UV-DDB-dependent damage recognition pathway predominates over direct recognition by XPC, and they also suggest the presence of rate-limiting regulatory steps in NER prior to the assembly of XPA.

Original languageEnglish
Pages (from-to)767-776
Number of pages10
JournalDNA Repair
Volume8
Issue number6
DOIs
Publication statusPublished - 4 Jun 2009
Externally publishedYes

Keywords

  • Dynamics
  • Nucleotide excision repair
  • UV-DDB
  • XPC

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