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: A J van Vuuren, E Appeldoorn, H Odijk, A Yasui, N G Jaspers, D Bootsma, J H Hoeijmakers
• Journal: The EMBO journal (EMBO J) Vol. 12 Issue 9 Pg. 3693-701 (Sep 1993) ISSN: 0261-4189 [Print] England
• PMID: 8253091 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies
• DNA-Binding Proteins
• Fungal Proteins
• Proteins
• Saccharomyces cerevisiae Proteins
• xeroderma pigmentosum group F protein
• Acetoxyacetylaminofluorene
• ERCC1 protein, human
• Endonucleases
• RAD1 protein, S cerevisiae
• RAD10 protein, S cerevisiae
• Single-Strand Specific DNA and RNA Endonucleases
• DNA Repair Enzymes

Topics

• Acetoxyacetylaminofluorene (toxicity)
• Animals
• Antibodies (pharmacology)
• CHO Cells
• Cell-Free System
• Cockayne Syndrome (genetics)
• Cricetinae
• DNA Damage
• DNA Repair
• DNA Repair Enzymes
• DNA-Binding Proteins (metabolism)
• Endonucleases
• Fungal Proteins (metabolism)
• Genetic Complementation Test
• Humans
• Mutation
• Plasmids (drug effects, radiation effects)
• Protein Biosynthesis
• Proteins (genetics, immunology, metabolism)
• Saccharomyces cerevisiae (metabolism)
• Saccharomyces cerevisiae Proteins
• Single-Strand Specific DNA and RNA Endonucleases
• Transfection
• Ultraviolet Rays
• Xeroderma Pigmentosum (genetics, metabolism)