The Ogura sterility-inducing protein forms a large complex without interfering with the oxidative phosphorylation components in rapeseed mitochondria
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The Ogura cytoplasmic male sterility causing protein, ORF138, was found to be part of a complex with an apparent size of over 750 kDa in the inner membrane of mitochondria of sterile plants. ORF138 did not colocalize with any of the oxidative phosphorylation complexes, nor did its presence modify their apparent size or amount, compared to samples from fertile isogenic plants. We attempted to detect potential proteins or nucleic acids that could be involved in the large ORF138 complex by 2D PAGE, immunoprecipitation and nuclease treatments of native extracts. All our results suggest that the ORF138 protein is the main, if not only, component of this large complex. The capacities of complexes I, II, IV, and ATP synthase were identical in samples from sterile and fertile plants. Isolated mitochondria from sterile plants showed a higher oxygen consumption than those from fertile plants. In vivo respiration measurements suggest that the difference in O2 consumption measured at the organelle level is compensated at the cell/tissue level, completely in leaves, but only partially in male reproductive organs.
KeywordsBlue native polyacrylamide gel electrophoresis Cytoplasmic male sterility Mitochondria Protein complex Oxidative phosphorylation
Blue native polyacrylamide gel electrophoresis
Cytoplasmic male sterility
nDodecyl ßD maltoside
Liquid chromatography tandem mass spectrometry
Tandem affinity purification
We thank A. Martin-Canadell for growing the many plants that provided excellent starting material for the production of mitochondrial extracts. We are grateful to G. Bonnard and H. Fromm for the generous gift of antibodies against CPN60 and PHB, respectively. We thank B. Valot and L. Negroni, from the proteomics platform at the IFR87, for their help with the LC-MS/MS analyses. We are very grateful to G. Tcherkez and P. Gauthier, from the metabolomic platform at IFR87, for allowing us to use the Licor device and for their advice and help with gas exchange measurements. We thank M. Gonneau and H. Mireau for helpful and stimulating discussions during this work, and their critical reading of the manuscript. YD was supported by the Plant Genetics Department of INRA and the CETIOM (Centre technique interprofessionnel des oléagineux métropolitains).
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