Transfection of the cloned human excision repair gene ERCC-1 to UV-sensitive CHO mutants only corrects the repair defect in complementation group-2 mutants

M. van Duin, J. H. Janssen, J. de Wit, J. H J Hoeijmakers, L. H. Thompson, D. Bootsma, A. Westerveld

Research output: Contribution to journalArticle

28 Scopus citations

Abstract

The human DNA-excision repair gene ERCC-1 is cloned by its ability to correct the excision-repair defect of the ultraviolet light- and mitomycin-C-sensitive CHO mutant cell line 43-3B. This mutant is assigned to complementation group 2 of the excision-repair-deficient CHO mutants. In order to establish whether the correlation by ERCC-1 is confined to CHO mutants of one complementation group, the cloned repair gene, present on cosmid 34-43, was transfected to representative cell lines of the 6 complementation groups that have been identified to date. Following transfection, mycophenolic acid was used to select for transferants expressing the dominant marker gene Ecogpt, also present on cosmid 34-43. Cotransfer of the ERCC-1 gene was shown by Southern blot analysis of DNA from pooled (500-2000 independent colonies) transformants of each mutant. UV survival and UV-induced UDS showed that only mutants belonging to complementation group 2 and no mutants of other groups were corrected by the ERCC-1 gene. This demonstrates that ERCC-1 does not provide an aspecific bypass of excision-repair defects in CHO mutants and supports the assumption that the complementation analysis is based on mutations in different repair genes.

Original languageEnglish (US)
Pages (from-to)123-130
Number of pages8
JournalMutation Research DNA Repair Reports
Volume193
Issue number2
DOIs
StatePublished - 1988
Externally publishedYes

Keywords

  • CHO mutants
  • DNA excision repair
  • DNA transfection
  • ERCC-1

ASJC Scopus subject areas

  • Medicine(all)

Fingerprint Dive into the research topics of 'Transfection of the cloned human excision repair gene ERCC-1 to UV-sensitive CHO mutants only corrects the repair defect in complementation group-2 mutants'. Together they form a unique fingerprint.

  • Cite this