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PMID: 8811092 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Mutational analysis of the human nucleotide excision repair gene ERCC1.

Nucleic acids research ·Vol. 24 ·No. 17 ·1996-09-01 ·Pages 3370-80

Sijbers AM, van der Spek PJ, Odijk H, van den Berg J, van Duin M, Westerveld A, Jaspers NG, Bootsma D, Hoeijmakers JH

Abstract

The human DNA repair protein ERCC1 resides in a complex together with the ERCC4, ERCC11 and XP-F correcting activities, thought to perform the 5' strand incision during nucleotide excision repair (NER). Its yeast counterpart, RAD1-RAD10, has an additional engagement in a mitotic recombination pathway, probably required for repair of DNA cross-links. Mutational analysis revealed that the poorly conserved N-terminal 91 amino acids of ERCC1 are dispensable for both repair functions, in contrast to a deletion of only four residues from the C-terminus. A database search revealed a strongly conserved motif in this C-terminus sharing sequence homology with many DNA break processing proteins, indicating that this part is primarily required for the presumed structure-specific endonuclease activity of ERCC1. Most missense mutations in the central region give rise to an unstable protein (complex). Accordingly, we found that free ERCC1 is very rapidly degraded, suggesting that protein-protein interactions provide stability. Survival experiments show that the removal of cross-links requires less ERCC1 than UV repair. This suggests that the ERCC1-dependent step in cross-link repair occurs outside the context of NER and provides an explanation for the phenotype of the human repair syndrome xeroderma pigmentosum group F.

MeSH Terms
Amino Acid Sequence Animals Cisplatin/pharmacology Conserved Sequence DNA Mutational Analysis DNA Repair/genetics DNA, Complementary/genetics DNA-Binding Proteins Dose-Response Relationship, Radiation Endonucleases/genetics Gene Amplification Genetic Complementation Test Humans Mitomycin/pharmacology Molecular Sequence Data Mutagenesis Mutagens/pharmacology Proteins/genetics Recombination, Genetic Rodentia/genetics Sequence Deletion Sequence Homology, Amino Acid Transfection Xeroderma Pigmentosum/etiology,genetics
Chemicals
DNA, Complementary DNA-Binding Proteins Mutagens Proteins Mitomycin ERCC1 protein, human Endonucleases Cisplatin
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Sijbers A M
Department of Cell Biology and Genetics, Erasmus University, Rotterdam, The Netherlands.
van der Spek P J
Odijk H
van den Berg J
van Duin M
Westerveld A
Jaspers N G
Bootsma D
Hoeijmakers J H
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/24.17.3370
Published
1996-09-01
Pages
3370-80
Language
English
Region
England
NLM ID
0411011
PMCID
PMC146110
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/24.17.3370 PMC Full Text View journal details
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