Carcinogenesis, Vol 18, 245-249, Copyright © 1997 by Oxford University Press
BC McKay, MA Francis and AJ Rainbow
We have previously reported the use of a recombinant nonreplicating
adenovirus type 5, Ad5HCMVsp1 lacZ, expressing the lacZ gene under control
of the human cytomegalovirus (HCMV) immediate early promoter to assess
repair of a UV-damaged reporter gene in UV and heat shock (HS) treated
cells. Heat shock and UV-enhanced reactivation (HSER and UVER) of
beta-galactosidase (beta-gal) activity for UV-irradiated Ad5HCMVsp1 lacZ in
normal human fibroblasts involved the transcription coupled repair (TCR)
pathway. However, this inducible DNA repair response was absent in p53
deficient tumour cell lines. In order to examine further the requirement
for p53 in HSER and UVER, we have examined host cell reactivation (HCR) of
the reporter construct in HS treated, UV treated and mock treated
Li-Fraumeni syndrome (LFS) fibroblasts, which are heterozygous for a p53
mutation, and immortalized LFS cell sublines, which express only mutant
p53. HCR of beta-gal activity for UV- irradiated Ad5HCMVsp1 lacZ was normal
in all LFS cells examined. However, HCR of beta-gal activity for
UV-irradiated Ad5HCMVsp1 lacZ was elevated by pretreatment of cells with
either UV or HS in normal diploid human fibroblasts, but not in LFS cells.
LFS cells appear to be deficient in an inducible pathway which stimulates
repair of the reporter gene. These results support a role for p53 in a HS
and UV inducible DNA repair response in human cells which is dependent on
TCR.
ARTICLES
Wildtype p53 is required for heat shock and ultraviolet light enhanced repair of a UV-damaged reporter gene
Department of Biology, McMaster University, Hamilton, Ontario, Canada.
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