Compartment-specific protection of iron-sulfur proteins by superoxide dismutase.
Iron and oxygen are essential but potentially toxic constituents of most organisms, and their transport is meticulously regulated both at the cellular and systemic levels. Compartmentalization may be a homeostatic mechanism for isolating these biological reactants in cells. To investigate this hypothesis, we have undertaken a genetic analysis of the interaction between iron and oxygen metabolism in Drosophila. We show that Drosophila iron regulatory protein-1 (IRP1) registers cytosolic iron and oxidative stress through its labile iron sulfur cluster by switching between cytosolic aconitase and RNA-binding functions. IRP1 is strongly activated by silencing and genetic mutation of the cytosolic superoxide dismutase (Sod1), but is unaffected by silencing of mitochondrial Sod2. Conversely, mitochondrial aconitase activity is relatively insensitive to loss of Sod1 function, but drops dramatically if Sod2 activity is impaired. This strongly suggests that the mitochondrial boundary limits the range of superoxide reactivity in vivo. We also find that exposure of adults to paraquat converts cytosolic aconitase to IRP1 but has no affect on mitochondrial aconitase, indicating that paraquat generates superoxide in the cytosol but not in mitochondria. Accordingly, we find that transgene-mediated overexpression of Sod2 neither enhances paraquat resistance in Sod1+ flies nor compensates for lack of SOD1 activity in Sod1-null mutants. We conclude that in vivo, superoxide is confined to the subcellular compartment in which it is formed, and that the mitochondrial and cytosolic SODs provide independent protection to compartment-specific protein iron-sulfur clusters against attack by superoxide generated under oxidative stress within those compartments.
Missirlis
Fanis
F
Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA.
Hu
Jianguo
J
Kirby
Kim
K
Hilliker
Arthur J
AJ
Rouault
Tracey A
TA
Phillips
John P
JP
D000154:Aconitate Hydratase / Q000737:chemistry; D000818:Animals; D002460:Cell Line; D003600:Cytosol / Q000201:enzymology / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004330:Drosophila; D006540:Herbicides; D007501:Iron / Q000378:metabolism; D035941:Iron Regulatory Protein 1 / Q000378:metabolism; D007506:Iron-Sulfur Proteins / Q000737:chemistry*; D008928:Mitochondria / Q000201:enzymology; D009154:Mutation; D018384:Oxidative Stress; D010100:Oxygen / Q000378:metabolism; D010269:Paraquat / Q000494:pharmacology; D011485:Protein Binding; D012313:RNA / Q000378:metabolism; D034622:RNA Interference; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000378:metabolism; D000072105:Superoxide Dismutase-1; D013481:Superoxides; D013997:Time Factors; D019076:Transgenes; D000099156:Superoxide Dismutase 2
D000154:Aconitate Hydratase; D018076:DNA, Complementary; D006540:Herbicides; D007501:Iron; D035941:Iron Regulatory Protein 1; D007506:Iron-Sulfur Proteins; D010100:Oxygen; D010269:Paraquat; D012313:RNA; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; D013481:Superoxides; D000099156:Superoxide Dismutase 2; C000606290:SOD1 protein, human