1. Sequential recruitment of the repair factors during NER: the role of XPG in initiating the resynthesis step
- Author
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Zhou Yajin, Thilo Riedl, Jean-Marc Egly, Marietta Y.W.T. Lee, Jean Philippe Lainé, Vincent Mocquet, Institut de génétique et biologie moléculaire et cellulaire (IGBMC), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Louis Pasteur - Strasbourg I, and Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
DNA Repair ,MESH: DNA Polymerase III ,0302 clinical medicine ,MESH: Endonucleases ,Replication Protein A ,MESH: DNA Repair ,chemistry.chemical_classification ,0303 health sciences ,biology ,General Neuroscience ,Nuclear Proteins ,MESH: Transcription Factors ,Cell biology ,DNA-Binding Proteins ,MESH: DNA Repair Enzymes ,030220 oncology & carcinogenesis ,PCNA complex ,Cyclin-Dependent Kinase Inhibitor p21 ,Ultraviolet Rays ,DNA repair ,DNA damage ,DNA-binding protein ,Article ,General Biochemistry, Genetics and Molecular Biology ,MESH: Cyclin-Dependent Kinase Inhibitor p21 ,Cell Line ,03 medical and health sciences ,Proliferating Cell Nuclear Antigen ,Humans ,MESH: Replication Protein A ,Molecular Biology ,Replication protein A ,DNA Polymerase III ,030304 developmental biology ,MESH: DNA Damage ,DNA ligase ,MESH: Humans ,General Immunology and Microbiology ,[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology ,Endonucleases ,Molecular biology ,MESH: Cell Line ,Proliferating cell nuclear antigen ,MESH: Hela Cells ,MESH: Proliferating Cell Nuclear Antigen ,DNA Repair Enzymes ,chemistry ,biology.protein ,MESH: Ultraviolet Rays ,MESH: Nuclear Proteins ,MESH: DNA-Binding Proteins ,DNA Damage ,HeLa Cells ,Transcription Factors ,Nucleotide excision repair - 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.
- Published
- 2007
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