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(Received for publication, July 31, 1997, and in revised form, August 27, 1997)
From the Department of Biological Sciences, Stanford University,
Stanford, California 94305
We have shown previously that Li-Fraumeni
syndrome fibroblasts homozygous for p53 mutations are deficient in the
removal of UV-induced cyclobutane pyrimidine dimers from genomic DNA,
but still proficient in the transcription-coupled repair pathway (Ford, J. M., and Hanawalt, P. C. (1995) Proc. Natl. Acad.
Sci. U. S. A. 92, 8876-8880). We have now utilized monoclonal
antibodies specific for cyclobutane pyrimidine dimers or 6-4 photoproducts, respectively, to measure their repair in UV-irradiated
human fibroblasts. Cells homozygous for p53 mutations were deficient in
the repair of both photoproducts, whereas cells heterozygous for mutant
p53 exhibited normal repair of 6-4 photoproducts, but decreased initial
rates of removal of cyclobutane pyrimidine dimers, compared with normal cells. The specificity of the effect of wild-type p53 on nucleotide excision repair was demonstrated in a p53 homozygous mutant cell line
containing a tetracycline-regulated wild-type p53 gene. Wild-type p53
expression and activity were suppressed in the presence of tetracycline, whereas withdrawal of tetracycline resulted in the induction of p53 expression, cell cycle checkpoint activation, and DNA
damage-induced apoptosis. The regulated expression of wild-type p53
resulted in the recovery of normal levels of repair of both cyclobutane
pyrimidine dimers and 6-4 photoproducts in genomic DNA, but did not
alter the transcription-coupled repair of cyclobutane pyrimidine
dimers. Therefore, the wild-type p53 gene product is an important
determinant of nucleotide excision repair activity in human cells.
Volume 272, Number 44,
Issue of October 31, 1997
pp. 28073-28080
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
Expression of Wild-type p53 Is Required for Efficient Global
Genomic Nucleotide Excision Repair in UV-irradiated Human
Fibroblasts
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