Gene-specific nuclear and mitochondrial repair of formamidopyrimidine DNA glycosylase-sensitive sites in Chinese hamster ovary cells.
This study examines the capacity of a mammalian cell to repair, at the gene level, DNA base lesions generated by photoactivation of acridine orange. Chinese hamster ovary fibroblasts were exposed to acridine orange and visible light, and gene-specific DNA repair was measured in the dihydrofolate reductase (DHFR) gene and in the mitochondrial genome. DNA lesions were recognized by Escherichia coli formamidepyrimidine-DNA glycosylase (FPG) which removes predominantly 8-oxodG and the corresponding formamidopyrimidine ring opened bases, and subsequently cleaves the DNA at the resulting apurinic site. FPG-recognized DNA lesions increased linearly with increasing photo-activation of AO, while cell survival was not affected by light alone and was negligibly affected by preincubation with AO in the dark. The frequency of induction of FPG-sensitive DNA damage by photoactivation of AO was similar in the transcribed and non-transcribed nuclear DNA as well as in the mitochondrial DNA. FPG-sensitive sites in the DHFR gene were repaired quickly, with 84% of adducts repaired within 4 h. The lesion frequency, kinetics and percent of repair of non-transcribed genomic DNA did not differ significantly from repair in the active DHFR gene up to 1 h postexposure. At late time points, transcribed DNA was repaired faster than the non-transcribed DNA. Mitochondrial DNA was efficiently repaired, at a rate similar to that in the active nuclear DNA.
Taffe
B G
BG
Wayne State University, Detroit, MI, USA.
Larminat
F
F
Laval
J
J
Croteau
D L
DL
Anson
R M
RM
Bohr
V A
VA
D000165:Acridine Orange / Q000494:pharmacology; D000818:Animals; D016466:CHO Cells / Q000187:drug effects; D002467:Cell Nucleus / Q000502:physiology; D002470:Cell Survival; D006224:Cricetinae; D004247:DNA / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D018736:DNA Adducts; D004260:DNA Repair / Q000235:genetics*; D004272:DNA, Mitochondrial / Q000187:drug effects / Q000235:genetics*; D043382:DNA-Formamidopyrimidine Glycosylase; D004926:Escherichia coli / Q000201:enzymology; D029968:Escherichia coli Proteins; D005796:Genes / Q000235:genetics*; D008027:Light; D009153:Mutagens / Q000494:pharmacology; D009699:N-Glycosyl Hydrolases / Q000378:metabolism*; D012045:Regulatory Sequences, Nucleic Acid / Q000187:drug effects; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics
D018736:DNA Adducts; D004272:DNA, Mitochondrial; D029968:Escherichia coli Proteins; D009153:Mutagens; D004247:DNA; D013762:Tetrahydrofolate Dehydrogenase; D009699:N-Glycosyl Hydrolases; D043382:DNA-Formamidopyrimidine Glycosylase; C468610:DNA-formamidopyrimidine glycosylase, E coli; D000165:Acridine Orange