title: Enhanced host cell reactivation of a UV-damaged reporter gene in pre-UV-treated cells is delayed in Cockayne syndrome cells.
abstract: We have used a non-replicating recombinant adenovirus, Ad5HCMVlacZ, which expresses the beta-galactosidase (beta-gal) reporter gene, to examine the time course of UV-inducible repair of UV-damaged DNA in human fibroblasts. Host cell reactivation (HCR) of beta-gal activity for UV-irradiated Ad5HCMVlacZ was examined in non-irradiated and UV-irradiated nucleotide excision repair (NER) proficient normal human fibroblasts, xeroderma pigmentosum (XP) group C fibroblasts which are defective in the global genomic repair (GGR) pathway of NER and Cockayne syndrome (CS) fibroblasts which are defective in the transcription coupled repair (TCR) pathway of NER. HCR was deficient in untreated XP-C and CS cells indicating that both TCR and GGR are involved in removal of photolesions from the transcribed strand of the reporter gene in unirradiated human cells as reported previously. Prior UV-irradiation of cells with low UV fluences resulted in a transient enhancement of HCR in normal and XP-C fibroblasts that reached a maximum when cells were infected at 25-35 h after UV. In contrast, UV-enhanced HCR was delayed in CS-B cells, reaching levels similar to that in normal cells only when cells were infected between 40 and 60 h after UV exposure. These results are consistent with a UV-induced up-regulation of GGR through a TCR dependent pathway in CS cells.
authors.affiliation: Department of Biology, McMaster University, 1280 Main Street West, Hamilton, Ont., Canada L8S 4K1.
authors.forename: Photini
authors.initials: P
authors.lastname: Pitsikas
authors.forename: Murray A
authors.initials: MA
authors.lastname: Francis
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Rainbow
mesh_terms: D003057:Cockayne Syndrome / Q000235:genetics* / Q000473:pathology; D004249:DNA Damage; D004260:DNA Repair; D005786:Gene Expression Regulation / Q000528:radiation effects*; D017930:Genes, Reporter; D006801:Humans; D014466:Ultraviolet Rays; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D001616:beta-Galactosidase