Nucleic Acids Research, Vol 25, Issue 14 2823-2827, Copyright © 1997 by Oxford University Press
C Rodel, T Jupitz and H Schmidt
In human cells DNA damage caused by UV light is mainly repaired by the
nucleotide excision repair pathway. This mechanism involves dual incisions
on both sides of the damage catalyzed by two nucleases. In mammalian cells
XPG cleaves 3' of the DNA lesion while the ERCC1-XPF complex makes the 5'
incision. The amino acid sequence of the human excision repair protein
ERCC1 is homologous with the fission yeast Swi10 protein. In order to test
whether these proteins are functional homologues, we overexpressed the
human gene in a Schizosaccharomyces pombe swi10 mutant. A swi10 mutation
has a pleiotropic effect: it reduces the frequency of mating type switching
(a mitotic transposition event from a silent cassette into the expression
site) and causes increased UV sensitivity. We found that the full-length
ERCC1 gene only complements the transposition defect of the fission yeast
mutant, while a C-terminal truncated ERCC1 protein also restores the DNA
repair capacity of the yeast cells. Using the two-hybrid system of
Saccharomyces cerevisiae we show that only the truncated human ERCC1
protein is able to interact with the S . pombe Rad16 protein, which is the
fission yeast homologue of human XPF. This is the first example yet known
that a human gene can correct a yeast mutation in nucleotide excision
repair.
ARTICLES
Complementation of the DNA repair-deficient swi10 mutant of fission yeast by the human ERCC1 gene
Institute of Genetics, Technische Universitat Braunschweig, Spielmannstrasse 7, D-38106 Braunschweig, Germany.
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