Oxidative stress impairs the repair of oxidative DNA base modifications in human skin fibroblasts and melanoma cells.
Irradiation of mammalian cells with solar light is associated with the generation of reactive oxygen species (ROS) and oxidative stress, which is mediated in part by endogenous photosensitizers absorbing in the visible range of the solar spectrum. Accordingly, oxidative DNA base modifications such as 7,8-dihydro-8-oxoguanine (8-oxoG) are the predominant types of DNA damage in cells irradiated at wavelengths >400 nm. We have analysed the repair of oxidative purine modifications in human skin fibroblasts and melanoma cells using an alkaline elution technique, both under normal conditions and after depletion of glutathione. Similar repair rates were observed in fibroblasts and melanoma cells from three different patients (t1/2 approximately 4h). In both cell types, glutathione depletion (increased oxidative stress) caused a pronounced repair retardation even under non-toxic irradiation conditions. Furthermore, the cleavage activity at 8-oxoG residues measured in protein extracts of the cells dropped transiently after irradiation. An addition of dithiothreitol restored normal repair rates. Interestingly, the repair rates of cyclobutane pyrimidine dimers (t1/2 approximately 18 h), AP sites (t1/2 approximately 1h) and single-strand breaks (t1/2 <30 min) were not affected by the light-induced oxidative stress. We conclude that the base excision repair of oxidative purine modifications is surprisingly vulnerable to oxidative stress, while the nucleotide excision repair of pyrimidine dimers is not.
Eiberger
Wolfgang
W
Institute of Pharmacy, University of Mainz, Staudingerweg 5, 55099 Mainz, Germany.
Volkmer
Beate
B
Amouroux
Rachel
R
Dhérin
Claudine
C
Radicella
J Pablo
JP
Epe
Bernd
B
D000368:Aged; D004260:DNA Repair; D005260:Female; D005978:Glutathione / Q000378:metabolism; D006151:Guanosine / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D008027:Light; D008545:Melanoma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D018384:Oxidative Stress; D012867:Skin / Q000166:cytology / Q000378:metabolism*; D014466:Ultraviolet Rays
D006151:Guanosine; C046215:8-hydroxyguanosine; D005978:Glutathione