title: Evidence for a repair enzyme complex involving ERCC1 and complementing activities of ERCC4, ERCC11 and xeroderma pigmentosum group F.
abstract: 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.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Cell Biology and Genetics, Erasmus University, Rotterdam, The Netherlands.
authors.forename: A J
authors.initials: AJ
authors.lastname: van Vuuren
authors.forename: E
authors.initials: E
authors.lastname: Appeldoorn
authors.forename: H
authors.initials: H
authors.lastname: Odijk
authors.forename: A
authors.initials: A
authors.lastname: Yasui
authors.forename: N G
authors.initials: NG
authors.lastname: Jaspers
authors.forename: D
authors.initials: D
authors.lastname: Bootsma
authors.forename: J H
authors.initials: JH
authors.lastname: Hoeijmakers
mesh_terms: D000099:Acetoxyacetylaminofluorene / Q000633:toxicity; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D016466:CHO Cells; D002474:Cell-Free System; D003057:Cockayne Syndrome / Q000235:genetics; D006224:Cricetinae; D004249:DNA Damage; D004260:DNA Repair; D045643:DNA Repair Enzymes; D004268:DNA-Binding Proteins / Q000378:metabolism; D004720:Endonucleases; D005656:Fungal Proteins / Q000378:metabolism; D005816:Genetic Complementation Test; D006801:Humans; D009154:Mutation; D010957:Plasmids / Q000187:drug effects / Q000528:radiation effects; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D015719:Single-Strand Specific DNA and RNA Endonucleases; D014162:Transfection; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum / Q000235:genetics* / Q000378:metabolism
chemical_list: D000906:Antibodies; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D011506:Proteins; D029701:Saccharomyces cerevisiae Proteins; C095888:xeroderma pigmentosum group F protein; D000099:Acetoxyacetylaminofluorene; C071447:ERCC1 protein, human; D004720:Endonucleases; C069266:RAD1 protein, S cerevisiae; C064162:RAD10 protein, S cerevisiae; D015719:Single-Strand Specific DNA and RNA Endonucleases; D045643:DNA Repair Enzymes