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PMID: 18079701 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Sequential recruitment of the repair factors during NER: the role of XPG in initiating the resynthesis step.

The EMBO journal ·Vol. 27 ·No. 1 ·2008-01-09 ·Pages 155-67

Mocquet V, Lainé JP, Riedl T, Yajin Z, Lee MY, Egly JM

Abstract

To address the biochemical mechanisms underlying the coordination between the various proteins required for nucleotide excision repair (NER), we employed the immobilized template system. Using either wild-type or mutated recombinant proteins, we identified the factors involved in the NER process and showed the sequential comings and goings of these factors to the immobilized damaged DNA. Firstly, we found that PCNA and RF-C arrival requires XPF 5' incision. Moreover, the positioning of RF-C is facilitated by RPA and induces XPF release. Concomitantly, XPG leads to PCNA recruitment and stabilization. Our data strongly suggest that this interaction with XPG protects PCNA and Pol delta from the effect of inhibitors such as p21. XPG and RPA are released as soon as Pol delta is recruited by the RF-C/PCNA complex. Finally, a ligation system composed of FEN1 and Ligase I can be recruited to fully restore the DNA. In addition, using XP or trichothiodystrophy patient-derived cell extracts, we were able to diagnose the biochemical defect that may prove to be important for therapeutic purposes.

MeSH Terms
Cell Line Cyclin-Dependent Kinase Inhibitor p21/physiology DNA Damage/physiology DNA Polymerase III/antagonists & inhibitors,metabolism DNA Repair/physiology DNA Repair Enzymes/metabolism DNA-Binding Proteins/physiology Endonucleases/physiology HeLa Cells Humans Nuclear Proteins/physiology Proliferating Cell Nuclear Antigen/metabolism Replication Protein A/metabolism Transcription Factors/physiology Ultraviolet Rays
Chemicals
CDKN1A protein, human Cyclin-Dependent Kinase Inhibitor p21 DNA excision repair protein ERCC-5 DNA-Binding Proteins Nuclear Proteins Proliferating Cell Nuclear Antigen Replication Protein A Transcription Factors DNA Polymerase III Endonucleases DNA Repair Enzymes
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Mocquet Vincent
Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch Cedex, France.
Lainé Jean Philippe
Riedl Thilo
Yajin Zhou
Lee Marietta Y
Egly Jean Marc
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
1460-2075
DOI
10.1038/sj.emboj.7601948
Published
2008-01-09
Epub
2007-00-13
Pages
155-67
Language
English
Region
England
NLM ID
8208664
PMCID
PMC2206130
Subset
IM
External Links
PubMed source DOI: 10.1038/sj.emboj.7601948 PMC Full Text View journal details
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