Mitochondrial Superoxide Decreases Yeast Survival in Stationary Phase
Longo VD, Liou L-L, Valentine JS, Gralla EB
Archives of Biochemistry and Biophysics (1999)
Category: iron-sulfur clusters, mitochondria, oxidative stress ¤ Added: Mar 16, 2006 ¤ Rating: ◊◊
Yeast lacking mitochondrial superoxide dismutase (MnSOD) display shortened stationary-phase survival and provide a good model system for studying mitochondrial oxidative damage. We observed a marked decrease in respiratory function preceding stationaryphase death of yeast lacking MnSOD (sod2D). Agents (mitochondrial inhibitors) that are known to increase or decrease superoxide production in submitochondrial particles affected stationary-phase survival in a manner inversely correlated with their effects on superoxide production, implicating superoxide in this mitochondrial disfunction. Similar but less-dramatic effects were observed in wild-type yeast. The activities of certain mitochondrial enzymes were particularly affected. In sod2D yeast the activity of aconitase, a 4Fe–4S-cluster-containing enzyme located in the matrix, was greatly and progressively decreased as the cells established stationary phase. Succinate dehydrogenase activity also decreased in MnSOD mutants; cytochrome oxidase and ATPase activities did not. Aconitase could be reactivated by addition of materials required for cluster assembly (Fe31 and a sulfur source), both in extracts and in vivo, indicating that inactivation of the enzyme was by disassembly of the cluster. Our results support the conclusion that superoxide is generated in the mitochondria in vivo and under physiological conditions and that MnSOD is the primary defense against this toxicity. When the balance between superoxide generation and MnSOD activity is disrupted, superoxide mediates iron release from mitochondrial iron–sulfur clusters, leading first to loss of mitochondrial function and then to death, independently of mtDNA damage. These results raise the possibility that similar processes may occur in higher eukaryotes. © 1999 Academic Press Key Words: manganese superoxide dismutase; SOD2; yeast; Saccharomyces cerevisiae; oxidative stress; mitochondria; aconitase; stationary phase.