A simultaneous release of SOD1 with cytochrome c regulates mitochondria-dependent apoptosis.
To elucidate the significance of mitochondrial localization of Cu/Zn-SOD (SOD1), we studied the relationship between the release of mitochondrial SOD1 and apoptosis. Kinetic analysis using HL-60 cells showed that both mitochondria-dependent and mitochondria-independent pro-apoptotic drugs, such as staurosporine and actinomycin D, increased the generation of reactive oxygen species (ROS) and decreased mitochondrial membrane potential (Delta psi). ROS generation by these drugs was inhibited by Mn (III) tetrakis (5,10,15,20-benzoic acid) porphyrin (MnTBAP), a cell membrane-permeable SOD mimetic. However, MnTBAP inhibited the apoptosis induced by staurosporine but not by actinomycin D. MnTBAP failed to inhibit Delta psi decrease and release of SOD1 and cytochrome c induced by actinomycin D. Moreover, 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), an inhibitor of voltage-dependent anion channel (VDAC), inhibited the release of the two proteins and apoptosis induced by staurosporine but not actinomycin D. These results suggest that ROS plays an important role in mitochondria-dependent but not mitochondria-independent apoptosis and that the release of SOD1 increases the susceptibility of mitochondria to oxidative stress, thereby enhancing a vicious cycle leading to apoptosis.
Li
Quan
Q
Department of Biochemistry and Molecular Pathology, Osaka City University Medical School, 1-4-3 Asahimachi, Abeno, Osaka, 545-8585, Japan.
Sato
Eisuke F
EF
Zhu
Xiaoping
X
Inoue
Masayasu
M
D017209:Apoptosis; D045304:Cytochromes c / Q000378:metabolism*; D053938:DNA Fragmentation; D003609:Dactinomycin / Q000494:pharmacology; D018922:HL-60 Cells; D006801:Humans; D053078:Membrane Potential, Mitochondrial; D008928:Mitochondria / Q000201:enzymology* / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D019311:Staurosporine / Q000494:pharmacology; D013482:Superoxide Dismutase / Q000378:metabolism*; D000072105:Superoxide Dismutase-1; D013997:Time Factors; D050994:Voltage-Dependent Anion Channels / Q000378:metabolism
D017382:Reactive Oxygen Species; C000606290:SOD1 protein, human; D050994:Voltage-Dependent Anion Channels; D003609:Dactinomycin; D045304:Cytochromes c; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; D019311:Staurosporine