The basal levels of 8-oxoG and other oxidative modifications in intact mitochondrial DNA are low even in repair-deficient (Ogg1(-/-)/Csb(-/-)) mice.
Mitochondrial DNA (mtDNA) is assumed to be highly prone to damage by reactive oxygen species (ROS) because of its location in close proximity to the mitochondrial electron transport chain. Accordingly, mitochondrial oxidative DNA damage has been hypothesized to be responsible for various neurological diseases, ageing and cancer. Since 7,8-dihydro-8-oxoguanine (8-oxoG), one of the most frequent oxidative base modifications, is removed from the mitochondrial genome by the glycosylase OGG1, the basal levels of this lesion are expected to be highly elevated in Ogg1(-/-) mice. To investigate this hypothesis, we have used a mtDNA relaxation assay in combination with various repair enzymes (Fpg, MutY, endonuclease III, endonuclease IV) to determine the average steady-state number of oxidative DNA modifications within intact (supercoiled) mtDNA from the livers of wild-type mice and those deficient in OGG1 and/or the Cockayne syndrome B (CSB) protein for mice aged up to 23 months. The levels of all types of oxidative modifications were found to be less than 12 per million base pairs, and the difference between wild-type and repair-deficient (Ogg1(-/-)/Csb(-/-)) mice was not significant. Thus, the increase of 8-oxoG caused by the repair deficiency in intact mtDNA is not much higher than in the nuclear DNA, i.e., not more than a few modifications per million base pairs. Based on these data, it is hypothesized that the load of oxidative base modifications in mtDNA is efficiently reduced during replication even in the absence of excision repair.
Institute of Pharmacy, University of Mainz, D-55099 Mainz, Germany.
Christian
C
Trapp
Amanda K
AK
McCullough
Bernd
B
Epe
D000818:Animals; D019240:Corticoviridae / Q000378:metabolism; D004249:DNA Damage; D045647:DNA Glycosylases / Q000172:deficiency* / Q000235:genetics; D004260:DNA Repair / Q000235:genetics / Q000502:physiology*; D045643:DNA Repair Enzymes / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D004272:DNA, Mitochondrial / Q000737:chemistry / Q000378:metabolism*; D004279:DNA, Viral / Q000737:chemistry / Q000378:metabolism; D005260:Female; D006151:Guanosine / Q000031:analogs & derivatives* / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010936:Plant Extracts / Q000378:metabolism; D000075223:Poly-ADP-Ribose Binding Proteins
D004272:DNA, Mitochondrial; D004279:DNA, Viral; C112288:LI 1370; D010936:Plant Extracts; D000075223:Poly-ADP-Ribose Binding Proteins; D006151:Guanosine; C046215:8-hydroxyguanosine; D045647:DNA Glycosylases; C485990:Ogg1 protein, mouse; C513075:Ercc6 protein, mouse; D045643:DNA Repair Enzymes