Co-correction of the ERCC1, ERCC4 and xeroderma pigmentosum group F DNA repair defects in vitro

M. Biggerstaff, D. E. Szymkowski, R. D. Wood

Research output: Contribution to journalArticle

135 Citations (Scopus)

Abstract

The mammalian ERCC1-encoded polypeptide is required for nucleotide excision repair of damaged DNA and is homologous to Saccharomyces cerevisiae RAD10, which functions in repair and mitotic intrachromosomal recombination. Rodent cells representing repair complementation group 1 have nonfunctional ERCC1. We report that repair of W-irradiated DNA can be reconstituted by combining rodent group 1 cell extracts with correcting protein from HeLa cells. Background repair was minimized by employing fractionated rodent cell extracts supplemented with human replication proteins RPA and PCNA. Group 1-correcting activity has a native molecular mass of 100 kDa and contains the 33 kDa ERCC1 polypeptide, as well as complementing activities for extracts from rodent group 4 and xeroderma pigmentosum group F (XP-F) cells. Extracts of group 1, group 4 or XP-F cells do not complement one another in vitro, although they complement extracts from other groups. The amount of ERCC1 detectable by immunoblotting is reduced in group 1, group 4 and XP-F extracts. Recombinant ERCC1 from Escherichia coli only weakly corrected the group 1 defect. The data suggest that ERCC1 is part of a functional protein complex with group 4 and XP-F correcting activities. The latter two may be equivalent to one another and analogous to S.cervisiae RAD1.

Original languageEnglish (US)
Pages (from-to)3685-3692
Number of pages8
JournalEMBO Journal
Volume12
Issue number9
StatePublished - Jan 1 1993

Fingerprint

Xeroderma Pigmentosum
DNA Repair
Rodentia
Repair
Defects
DNA
Cell Extracts
Peptides
Proteins
Proliferating Cell Nuclear Antigen
HeLa Cells
Immunoblotting
Genetic Recombination
Saccharomyces cerevisiae
Molecular mass
Yeast
Escherichia coli
Nucleotides
In Vitro Techniques

Keywords

  • Excision repair
  • Protein complex
  • RAD mutants
  • UV light
  • XP

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Co-correction of the ERCC1, ERCC4 and xeroderma pigmentosum group F DNA repair defects in vitro. / Biggerstaff, M.; Szymkowski, D. E.; Wood, R. D.

In: EMBO Journal, Vol. 12, No. 9, 01.01.1993, p. 3685-3692.

Research output: Contribution to journalArticle

Biggerstaff, M. ; Szymkowski, D. E. ; Wood, R. D. / Co-correction of the ERCC1, ERCC4 and xeroderma pigmentosum group F DNA repair defects in vitro. In: EMBO Journal. 1993 ; Vol. 12, No. 9. pp. 3685-3692.
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