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PMID: 3053162 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

A yeast DNA repair gene partially complements defective excision repair in mammalian cells.

The EMBO journal ·Vol. 7 ·No. 10 ·1988-10-00 ·Pages 3245-53

Lambert C, Couto LB, Weiss WA, Schultz RA, Thompson LH, Friedberg EC

Abstract

The RAD10 gene of Saccharomyces cerevisiae is required for nucleotide excision repair of DNA. Expression of RAD10 mRNA and Rad10 protein was demonstrated in Chinese hamster ovary (CHO) cells containing amplified copies of the gene, and RAD10 mRNA was also detected in stable transfectants without gene amplification. Following transfection with the RAD10 gene, three independently isolated excision repair-defective CHO cell lines from the same genetic complementation group (complementation group 2) showed partial complementation of sensitivity to killing by UV radiation and to the DNA cross-linking agent mitomycin C. These results were not observed when RAD10 was introduced into excision repair-defective CHO cell lines from other genetic complementation groups, nor when the yeast RAD3 gene was expressed in cells from genetic complementation group 2. Enhanced UV resistance in cells carrying the RAD10 gene was accompanied by partial reactivation of the plasmid-borne chloramphenicol acetyltransferase (cat) gene following its inactivation by UV radiation. The phenotype of CHO cells from genetic complementation group 2 is also specifically complemented by the human ERCC1 gene, and the ERCC1 and RAD10 genes have similar amino acid sequences. The present experiments therefore indicate that the structural homology between the yeast Rad10 and human Ercc1 polypeptides is reflected at a functional level, and suggest that nucleotide excision repair proteins are conserved in eukaryotes.

MeSH Terms
Animals Blotting, Northern Blotting, Western Cell Line Cricetinae DNA Repair DNA, Recombinant Gene Expression Regulation Genes, Fungal Genetic Complementation Test Nuclear Proteins/genetics Phenotype RNA, Messenger/genetics Saccharomyces cerevisiae/genetics
Chemicals
DNA, Recombinant Nuclear Proteins RNA, Messenger
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Lambert C
Department of Pathology, Stanford University School of Medicine, CA 94305.
Couto L B
Weiss W A
Schultz R A
Thompson L H
Friedberg E C
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1988-10-00
Pages
3245-53
Language
English
Region
England
NLM ID
8208664
PMCID
PMC454734
Subset
IM
Grants
NCI NIH HHS · CA-07705 · United States
NCI NIH HHS · CA-09302 · United States
NCI NIH HHS · CA-12428 · United States
External Links
PubMed source PMC Full Text View journal details
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