Radiation-induced gene expression was examined in cells of a radioresistant human glioblastoma cell line, T98G, using RNA fingerprinting by arbitrary primer polymerase chain reaction. Three of the differentially induced transcripts were cloned and identified as the mitochondrially encoded cytochrome c oxidase (MTCO) subunits 1 and 2, and NADH dehydrogenase subunit 4. The levels of all three mRNAs were increased 1 h after irradiation, with elevated expression persisting for at least 24 h. Similar to radiation, other oxidants lead to induction of MTCO1, an effect which could be prevented by the antioxidant pyrrolidine dithiocarbamate. These results indicate that the increase in mitochondrial gene expression is mediated by oxidative stress. Mitochondria could be a target of signaling by ionizing radiation and oxidants since it is known to be at the site of cellular oxidative stress. The proteins encoded by MTCO1 and other mitochondrial mRNAs characterized are part of the mitochondrial respiratory chain which produces adenosine triphosphate through the process of oxidative phosphorylation. Adenosine triphosphate levels and the mitochondrial membrane potential were found to be increased significantly after irradiation, while mitochondrial mass and mitochondrial DNA levels were unaffected. These data demonstrate the specificity of changes in mitochondrial activity and gene expression after irradiation.