Evidence for a repair enzyme complex involving ERCC1, and the correcting activities of ERCC4, ERCC11 and the xeroderma pigmentosum group F.
Nucleotide excision repair (NER), one of the major cellular DNA repair systems, removes a wide range of lesions in a multi-enzyme reaction. In man, a NER defect due to a mutation in one of at least 11 distinct genes, can give rise to the inherited repair disorders xeroderma pigmentosum (XP), Cockayne's syndrome or PIBIDS, a photosensitive form of the brittle hair disease trichothiodystrophy. Laboratory-induced NER-deficient mutants of cultured rodent cells have been classified into 11 complementation groups (CGs). Some of these have been shown to correspond with human disorders. In cell-free extracts prepared from rodent CGs 1-5 and 11, but not in a mutant from CG6, we find an impaired repair of damage induced in plasmids by UV light and N-acetoxy-acetylaminofluorene. Complementation analysis in vitro of rodent CGs is accomplished by pairwise mixing of mutant extracts. The results show that mutants from groups 2, 3, 5 and XP-A can complement all other CGs tested. However, selective non-complementation in vitro was observed in mutual mixtures of groups 1, 4, 11 and XP-F, suggesting that the complementing activities involved somehow affect each other. Depletion of wild-type human extracts from ERCC1 protein using specific anti-ERCC1 antibodies concomitantly removed the correcting activities for groups 4, 11 and XP-F, but not those for the other CGs. Furthermore, we find that 33 kDa ERCC1 protein sediments as a high mol. wt species of approximately 120 kDa in a native glycerol gradient.
|Keywords||*DNA Damage, *DNA Repair, *Endonucleases, 0 (Antibodies), 0 (DNA-Binding Proteins), 0 (ERCC4 protein), 0 (Fungal Proteins), 0 (Plasmids), 0 (Proteins), 0 (RAD1 protein, S cerevisiae), 6098-44-8 (Acetoxyacetylaminofluorene), Acetoxyacetylaminofluorene/toxicity, Animals, Antibodies/pharmacology, CHO Cells, Cell-Free System, Cockayne Syndrome/genetics, Comparative Study, DNA-Binding Proteins/metabolism, EC 3.1.- (ERCC-1 protein, human), EC 3.1.- (Endonucleases), EC 3.1.- (RAD10 protein, S cerevisiae), Fungal Proteins/metabolism, Genetic Complementation Test, Hamsters, Human, Mutation, Plasmids/drug effects/radiation effects, Proteins/genetics/immunology/*metabolism, Saccharomyces cerevisiae/metabolism, Support, Non-U.S. Gov't, Transfection, Translation, Genetic, Ultraviolet Rays, Xeroderma Pigmentosum/*genetics/metabolism|
van Vuuren, A.J., Appeldoorn, E., Odijk, H., Yasui, A., Jaspers, N.G.J., Bootsma, D., & Hoeijmakers, J.H.J.. (1993). Evidence for a repair enzyme complex involving ERCC1, and the correcting activities of ERCC4, ERCC11 and the xeroderma pigmentosum group F.. EMBO Journal, 12, 3693–3701. Retrieved from http://hdl.handle.net/1765/3055