Wildtype p53 is required for heat shock and ultraviolet light enhanced repair of a UV-damaged reporter gene.
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.
McKay
B C
BC
Department of Biology, McMaster University, Hamilton, Ontario, Canada.
Francis
M A
MA
Rainbow
A J
AJ
D000256:Adenoviridae / Q000235:genetics; D002460:Cell Line; D002470:Cell Survival; D004260:DNA Repair / Q000235:genetics* / Q000528:radiation effects; D004789:Enzyme Activation / Q000528:radiation effects; D016158:Genes, p53 / Q000502:physiology*; D006358:Hot Temperature; D006801:Humans; D007763:Lac Operon / Q000235:genetics / Q000528:radiation effects; D016864:Li-Fraumeni Syndrome / Q000201:enzymology / Q000235:genetics*; D014466:Ultraviolet Rays; D001616:beta-Galactosidase / Q000378:metabolism*
D001616:beta-Galactosidase