Evidence for a repair enzyme complex involving ERCC1 and complementing activities of ERCC4, ERCC11 and xeroderma pigmentosum group F

EMBO J. 1993 Sep;12(9):3693-701.

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)

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

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

Substances

  • 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