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V-ATPase activity in the TGN/EE is required for exocytosis and recycling in Arabidopsis

Nature Plants volume 1, Article number: 15094 (2015) Cite this article

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Abstract

In plants, vacuolar H+-ATPase (V-ATPase) activity acidifies both the trans-Golgi network/early endosome (TGN/EE) and the vacuole. This dual V-ATPase function has impeded our understanding of how the pH homeostasis within the plant TGN/EE controls exo- and endocytosis. Here, we show that the weak V-ATPase mutant deetiolated3 (det3) displayed a pH increase in the TGN/EE, but not in the vacuole, strongly impairing secretion and recycling of the brassinosteroid receptor and the cellulose synthase complexes to the plasma membrane, in contrast to mutants lacking tonoplast-localized V-ATPase activity only. The brassinosteroid insensitivity and the cellulose deficiency defects in det3 were tightly correlated with reduced Golgi and TGN/EE motility. Thus, our results provide strong evidence that acidification of the TGN/EE, but not of the vacuole, is indispensable for functional secretion and recycling in plants.

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Figure 1: pH in the TGN/EE of det3 is increased.
Figure 2: Reduced Golgi and TGN/EE motilities in det3.
Figure 3: Endocytic trafficking from TGN/EE to the vacuole is delayed in det3.
Figure 4: Secretion of BRI1 in det3 is reduced.
Figure 5: BRI1 recycling from TGN/EE to PM is reduced in det3.
Figure 6: Delivery of CesA complexes to the PM is impaired in det3.

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Acknowledgements

We thank Yanhai Yin (Iowa State University, Ames, USA) for providing the anti-BES1 antibody, Niko Geldner (Université de Lausanne, Switzerland) for sharing pBRI1:BRI1-GFP and pHS:BRI1-YFP lines, Daniël Van Damme (VIB-Ghent University, Belgium) for advice in microscopy, and Martine De Cock (Ghent University, Belgium) for help in preparing the manuscript. This work was supported by the Marie Curie Initial Training Network ‘BRAVISSIMO’ (grant no. PITN-GA-2008-215118) to E.R., the Odysseus program of the Research Foundation-Flanders to J.F., the Max-Planck Gesellschaft to A.D., Y.Z., L.N. and S.P., the European Union Seventh Framework Programme (FP7 2007–2013) under Grant Agreement 263916 (WallTrac, Marie Curie Initial Training Network) to P.K. and the Deutsche Forschungsgemeinschaft (SFB1101 and TPA02) to K.S. Y.L. is indebted to the Belgian Science Policy Office (BELSPO) for a postdoctoral fellowship.

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Author notes

  1. Niloufer G. Irani

    Present address: ‡Present address: Department of Plant Sciences, University of Oxford, Oxford OX1 3RB, UK,

  2. Yu Luo, Stefan Scholl, Anett Doering and Staffan Persson: These authors contributed equally to this work

  3. Karin Schumacher and Eugenia Russinova: These authors jointly supervised this work.

Authors and Affiliations

  1. Department of Plant Systems Biology, VIB, Gent, 9052, Belgium

    Yu Luo, Niloufer G. Irani, Simone Di Rubbo, Isabelle Van Houtte, Evelien Mylle & Eugenia Russinova

  2. Department of Plant Biotechnology and Bioinformatics, Ghent University, Gent, 9052, Belgium

    Yu Luo, Niloufer G. Irani, Simone Di Rubbo, Isabelle Van Houtte, Evelien Mylle & Eugenia Russinova

  3. Developmental Biology of Plants, Centre for Organismal Studies (COS), Heidelberg University, Heidelberg, 69120, Germany

    Stefan Scholl & Karin Schumacher

  4. Max-Planck Institute for Molecular Plant Physiology, Potsdam, 14476, Germany

    Anett Doering, Yi Zhang, Lutz Neumetzler, Praveen Krishnamoorthy & Staffan Persson

  5. Institut National de la Recherche Agronomique, Unité Mixte de Recherche 1318, Institut Jean-Pierre Bourgin, Saclay Plant Sciences, Versailles, 78000, France

    Volker Bischoff & Samantha Vernhettes

  6. AgroParisTech, Institut Jean-Pierre Bourgin, Versailles, 78000, France

    Volker Bischoff & Samantha Vernhettes

  7. Department of Organic Chemistry, Polymer Chemistry Research Group and Laboratory for Organic Synthesis, Ghent University, Gent, 9000, Belgium

    Johan Winne

  8. Institute of Science and Technology Austria (IST Austria), Klosterneuburg, 3400, Austria

    Jiří Friml

  9. Center for Plant Molecular Biology (ZMBP), University of Tübingen, Tübingen, 72076, Germany

    York-Dieter Stierhof

  10. Australian Research Council, Centre of Excellence in Plant Cell Walls, School of Biosciences, University of Melbourne, Parkville, Victoria, 3010, Australia

    Staffan Persson

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Contributions

Y.L., S.S., A.D., N.G.I., L.N., J.F, K.S., S.P. and E.R. conceived the study and designed the experiments. Y.L. performed all BR-related work, A.D., Y.Z. and P.K. performed all CesA-related work, S.S. constructed the pH sensor and did all pH measurements, N.G.I. did the AFCS uptake, Y.L., A.D., Y.Z., N.G.I., S.D.R. and I.V.H. generated materials, Y.-D. S. and K.S. did the TEM, Y.L., A.D., Y.Z., N.G.I., L.N., P.K., E.M., and V.B. did imaging, J.W. performed the chemical synthesis of AFCS, Y.L., A.D., N.G.I., S.D.R., K.S., S.P. and E.R. wrote the manuscript. All authors commented on the results and the manuscript.

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Correspondence to Karin Schumacher, Staffan Persson or Eugenia Russinova.

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Luo, Y., Scholl, S., Doering, A. et al. V-ATPase activity in the TGN/EE is required for exocytosis and recycling in Arabidopsis. Nature Plants 1, 15094 (2015). https://doi.org/10.1038/nplants.2015.94

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