The Ogura sterility-inducing protein forms a large complex without interfering with the oxidative phosphorylation components in rapeseed mitochondria
- 296 Downloads
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).
- Duroc Y, Gaillard C, Hiard S, Defrance MC, Pelletier G, Budar F (2005) Biochemical and functional characterization of ORF138, a mitochondrial protein responsible for Ogura cytoplasmic male sterility in Brassiceae. Biochimie 87:1089–1100. doi: 10.1016/j.biochi.2005.05.009 PubMedCrossRefGoogle Scholar
- Gonzalez-Melendi P, Uyttewaal M, Morcillo CN, Hernandez Mora JR, Fajardo S, Budar F, Lucas MM (2008) A light and electron microscopy analysis of the events leading to male sterility in Ogu-INRA CMS of rapeseed (Brassica napus). J Exp Bot 59:827–838. doi: 10.1093/jxb/erm365 PubMedCrossRefGoogle Scholar
- Juszczuk IM, Flexas J, Szal B, Dabrowska Z, Ribas-Carbo M, Rychter AM (2007) Effect of mitochondrial genome rearrangement on respiratory activity, photosynthesis, photorespiration and energy status of MSC16 cucumber (Cucumis sativus) mutant. Physiol Plant 131:527–541. doi: 10.1111/j.1399-3054.2007.00984.x PubMedCrossRefGoogle Scholar
- Priault P, Tcherkez G, Cornic G, De Paepe R, Naik R, Ghashghaie J, Streb P (2006) The lack of mitochondrial complex I in a CMSII mutant of Nicotiana sylvestris increases photorespiration through an increased internal resistance to CO2 diffusion. J Exp Bot 57:3195–3207. doi: 10.1093/jxb/erl083 PubMedCrossRefGoogle Scholar
- Rohila JS, Chen M, Chen S, Chen J, Cerny R, Dardick C, Canlas P, Xu X, Gribskov M, Kanrar S, Zhu JK, Ronald P, Fromm ME (2006) Protein–protein interactions of tandem affinity purification-tagged protein kinases in rice. Plant J 46:1–13. doi: 10.1111/j.1365-313X.2006.02671.x PubMedCrossRefGoogle Scholar
- Van Der Hoorn RA, Rivas S, Wulff BB, Jones JD, Joosten MH (2003) Rapid migration in gel filtration of the Cf-4 and Cf-9 resistance proteins is an intrinsic property of Cf proteins and not because of their association with high-molecular-weight proteins. Plant J 35:305–315. doi: 10.1046/j.1365-313X.2003.01803.x CrossRefGoogle Scholar