Mitochondrial GPx1 decreases induced but not basal oxidative damage to mtDNA in T47D cells.
The production of oxyradicals by mitochondria (mt) is a source of oxidative damage to mtDNA such as 8-oxo-dG lesions that may lead to mutations and mitochondrial dysfunction. The potential protection of mtDNA by glutathione peroxidase-1 (GPx1) was investigated in GPx1-proficient (GPx-2) and GPx1-deficient (Hygro-3) human breast T47D cell transfectants. GPx activity and GPx1-like antigen concentration in mitochondria were respectively at least 100-fold and 20- to 25-fold higher in GPx2 than Hygro-3 cells. In spite of this large difference in peroxide-scavenging capacity, the basal 8-oxo-dG frequency in mtDNA, assessed by carefully controlled postlabeling assay, was strikingly similar in both cell lines. In contrast, in response to menadione-mediated oxidative stress, induction of 8-oxo-dG and DNA strand breaks was much lower in the GPx1-proficient mitochondria (e.g., +14% 8-oxo-dG versus +54% in Hygro-3 after 1-h exposure to 25 microM menadione, P < 0.05). Our data indicate that the mitochondrial glutathione/GPx1 system protected mtDNA against damage induced by oxidative stress, but did not prevent basal oxidative damage to mtDNA, which, surprisingly, appeared independent of GPx1 status in the T47D model.
Legault
J
J
Unit of Health and Environment, CHUL Research Center and Laval University, Québec, Canada.
Carrier
C
C
Petrov
P
P
Renard
P
P
Remacle
J
J
Mirault
M E
ME
D000080242:8-Hydroxy-2'-Deoxyguanosine; D016477:Artifacts; D003593:Cytoplasm / Q000201:enzymology / Q000648:ultrastructure; D004249:DNA Damage / Q000187:drug effects / Q000235:genetics*; D004272:DNA, Mitochondrial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000032:analysis / Q000235:genetics; D005978:Glutathione / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000172:deficiency / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D016253:Microscopy, Immunoelectron; D008928:Mitochondria / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D018384:Oxidative Stress / Q000187:drug effects; D014162:Transfection; D014407:Tumor Cells, Cultured; D014812:Vitamin K / Q000494:pharmacology; D000094765:Glutathione Peroxidase GPX1
D004272:DNA, Mitochondrial; D014812:Vitamin K; D000080242:8-Hydroxy-2'-Deoxyguanosine; D005979:Glutathione Peroxidase; D003849:Deoxyguanosine; D005978:Glutathione; D000094765:Glutathione Peroxidase GPX1; C000722067:GPX1 protein, human