Repair of oxidative damage in nuclear DNA sequences with different transcriptional activities.
This study was designed to investigate the repair of oxidative damage in nuclear DNA sequences with different transcriptional activities. Chinese hamster ovary (CHO) cells were treated with the oxygen radical generator hypoxanthine/xanthine oxidase (Hyp/XO). Damage and repair were evaluated in 14-kb restriction fragments containing either the DHFR gene, a 3'-non-transcribed flanking region, or the c-fos gene using a quantitative Southern blot technique. Damage to the sugar-phosphate backbone and abasic sites were detected by measuring their lability in alkali conditions. Lesions in DNA bases were identified using the bacterial repair enzyme endonuclease III, which predominantly recognizes damage to thymines and cytosines, and formamidopyrimidine-DNA glycosylase, which recognizes 8-oxoguanine and purines with fractured imidazole rings. The results showed that similar amounts of all types of oxidative damage were produced in both the transcribed and non-transcribed sequences following a 1-h exposure to the radical generator. Repair in all sequences was rapid, with approximately 60% removal of lesions observed by 1 h. Therefore, within these sequences, the repair of oxidative lesions is much faster than that of other types of damage, such as those induced by alkylating toxins and UV irradiation, and the repair is not affected appreciably by transcriptional status.
Grishko
V I
VI
Department of Structural and Cellular Biology, University of South Alabama, Mobile 36688, USA.
Driggers
W J
WJ
LeDoux
S P
SP
Wilson
G L
GL
D000818:Animals; D016466:CHO Cells / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects / Q000235:genetics*; D002470:Cell Survival / Q000187:drug effects; D006224:Cricetinae; D003596:Cytosine / Q000378:metabolism; D004249:DNA Damage / Q000235:genetics; D004260:DNA Repair / Q000235:genetics*; D043382:DNA-Formamidopyrimidine Glycosylase; D043224:Deoxyribonuclease (Pyrimidine Dimer); D004305:Dose-Response Relationship, Drug; D004706:Endodeoxyribonucleases / Q000378:metabolism; D029968:Escherichia coli Proteins; D016762:Genes, fos; D019271:Hypoxanthine / Q000494:pharmacology; D009699:N-Glycosyl Hydrolases / Q000378:metabolism; D010084:Oxidation-Reduction; D013379:Substrate Specificity; D013762:Tetrahydrofolate Dehydrogenase / Q000187:drug effects / Q000235:genetics*; D013941:Thymine / Q000378:metabolism; D014158:Transcription, Genetic; D014969:Xanthine Oxidase / Q000494:pharmacology
D029968:Escherichia coli Proteins; D019271:Hypoxanthine; D003596:Cytosine; D014969:Xanthine Oxidase; D013762:Tetrahydrofolate Dehydrogenase; D004706:Endodeoxyribonucleases; D043224:Deoxyribonuclease (Pyrimidine Dimer); C106403:NTH protein, E coli; D009699:N-Glycosyl Hydrolases; D043382:DNA-Formamidopyrimidine Glycosylase; D013941:Thymine