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The Ogura sterility-inducing protein forms a large complex without interfering with the oxidative phosphorylation components in rapeseed mitochondria

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Abstract

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.

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Abbreviations

BN-PAGE:

Blue native polyacrylamide gel electrophoresis

CMS:

Cytoplasmic male sterility

CPN60:

Chaperone 60

DDM:

nDodecyl ßD maltoside

LC-MS/MS:

Liquid chromatography tandem mass spectrometry

PHB:

Prohibitin

PMSF:

Phenylmethanesulphonylfluoride

TAP:

Tandem affinity purification

nPG:

n-Propylgallate

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Acknowledgments

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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  1. Yann Duroc

    Present address: UPR2355, Centre National de la Recherche Scientifique, Bâtiment 23, 1 Avenue de la Terrasse, 91198, Gif-Sur-Yvette Cedex, France

  2. Sandrine Ragu

    Present address: UMR 2027 CNRS/Institut Curie, Batiment 110, Centre Universitaire, 91405, Orsay, France

Authors and Affiliations

  1. Station de Génétique et d’Amélioration des Plantes, Institut Jean-Pierre Bourgin, INRA UR254, Route de Saint-Cyr, 78026, Versailles cedex, France

    Yann Duroc, Sophie Hiard, Nathalie Vrielynck, Sandrine Ragu & Françoise Budar

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  1. Yann Duroc
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  2. Sophie Hiard
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Correspondence to Françoise Budar.

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S. Hiard and N. Vrielynck contributed equally to this work.

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Duroc, Y., Hiard, S., Vrielynck, N. et al. The Ogura sterility-inducing protein forms a large complex without interfering with the oxidative phosphorylation components in rapeseed mitochondria. Plant Mol Biol 70, 123–137 (2009). https://doi.org/10.1007/s11103-009-9461-6

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