Multiple C2 domains and Transmembrane region Proteins (MCTPs) tether membranes at plasmodesmata
- Creators
- Brault, Marie L.
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Petit, Jules D.
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Immel, Françoise
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Nicolas, William J.
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Glavier, Marie
- Brocard, Lysiane
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Gaston, Amélia
- Fouché, Mathieu
- Hawkins, Timothy J.
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Crowet, Jean-Marc
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Grison, Magali S.
- Germain, Véronique
- Rocher, Marion
- Kraner, Max
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Alva, Vikram
- Claverol, Stéphane
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Paterlini, Andrea
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Helariutta, Ykä
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Deleu, Magali
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Lins, Laurence
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Tilsner, Jens
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Bayer, Emmanuelle E.
Abstract
In eukaryotes, membrane contact sites (MCS) allow direct communication between organelles. Plants have evolved a unique type of MCS, inside intercellular pores, the plasmodesmata, where endoplasmic reticulum (ER)–plasma membrane (PM) contacts coincide with regulation of cell‐to‐cell signalling. The molecular mechanism and function of membrane tethering within plasmodesmata remain unknown. Here, we show that the multiple C2 domains and transmembrane region protein (MCTP) family, key regulators of cell‐to‐cell signalling in plants, act as ER‐PM tethers specifically at plasmodesmata. We report that MCTPs are plasmodesmata proteins that insert into the ER via their transmembrane region while their C2 domains dock to the PM through interaction with anionic phospholipids. A Atmctp3/Atmctp4 loss of function mutant induces plant developmental defects, impaired plasmodesmata function and composition, while MCTP4 expression in a yeast Δtether mutant partially restores ER‐PM tethering. Our data suggest that MCTPs are unique membrane tethers controlling both ER‐PM contacts and cell‐to‐cell signalling.
Additional Information
© 2019 The Authors. Published under the terms of the CC BY 4.0 license. Received 3 October 2018; Revised 28 May 2019; Accepted 6 June 2019; Published online 9 July 2019. Data availability: Proteomic data: PRIDE PXD006800 (http://www.ebi.ac.uk/pride/archive/projects/PXD006800), PRIDE PXD006806 (http://www.ebi.ac.uk/pride/archive/projects/PXD006806) and PRIDE PXD013999 (http://www.ebi.ac.uk/pride/archive/projects/PXD013999). This work was supported by the National Agency for Research (Grant ANR‐14‐CE19‐0006‐01 to E.M.B), "Osez l'interdisciplinarité" OSEZ‐2017‐BRIDGING CNRS programme to E.M.B., the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement No 772103‐BRIDGING) to E.M.B, the EMBO Young Investigator Program to E.M.B., and Fonds National de la Recherche Scientifique (NEAMEMB PDR T.1003.14, BRIDGING CDR J.0114.18 and RHAMEMB CDR J.0086.18) to L. L. and M.D. J.D.P. is funded by a PhD fellowship from the Belgian "Formation à la Recherche dans l'Industrie et l'Agriculture" (FRIA grant no. 1.E.096.18). Work in J.T. laboratory is supported by grant BB/M007200/1 from the U.K. Biotechnology and Biomedical Sciences Research Council (BBSRC) and by the Scottish Government's Rural and Environment Science and Analytical Services Division (RESAS). Fluorescence microscopy analyses were performed at the plant pole of the Bordeaux Imaging Centre (http://www.bic.u-bordeaux.fr). For electron microscopy, the Region Aquitaine also supported the acquisition of the electron microscope (grant no. 2011 13 04 007 PFM), and FranceBioImaging Infrastructure supported the acquisition of the AFS2 and ultramicrotome. The proteomic analyses were performed at the Functional Genomic Center of Bordeaux, (https://proteome.cgfb.u-bordeaux.fr). We thank Steffen Vanneste and Abel Rosado for providing the VAP27.1.RFP and SYT1.GFP binary vectors and Yvon Jaillais for providing the 1xPH(FAPP1) Arabidopsis transgenic lines. The plasmid pRBbar‐OCS was kindly provided by Prof. Frederik Börnke (IGZ—Leibniz Institute of Vegetable and Ornamental Crops, Groβbeeren, Germany). We thank Christophe Trehin and Patrice Morel for providing the AtMCT15_C2s construct and Alenka Copic for providing the yeast WT and Δtether strains. We thank Fabrice Cordelières for his help for the fluorescence image quantification and Paul Gouget, Yvon Jaillais, Andrea Paterlini and Yrjo Helariutta for critical review of the article prior to submission. Author contributions: FI, MSG, MF and SC carried out the label‐free proteomic analysis of plasmodesmata fractions isolated from Arabidopsis cell cultures. MLB cloned the MCTPs, produced and phenotyped the Arabidopsis transgenic lines, with the exception of AtMCTP4:GFP‐AtMCTP4 and 35S:GFP‐AtMCTP6 which were generated by MK. MLB and JDP imaged the MCTP reporter lines. WJN carried out the FRAP analysis and image quantification for co‐localisation with the help of LB. MG and JDP performed the CLEM and immunogold labelling approaches. JT, MR and VG performed the plasmodesmata cell‐to‐cell trafficking assay. MK carried out the proteomic comparison and AP the data integration. AG performed the phylogenic analysis. JDP and MLB carried out the PAO experiments. MLB performed the yeast experiments. TJH and JT performed the 3D‐SIM. VA carried out the C2 cluster map analysis. JDP carried out the molecular dynamic analysis with the help of J‐MC, LL and MD. EMB conceived the study and designed experiments with the help of JT and LL. EMB, JDP, JT, MLB and YH wrote the manuscript. All the authors discussed the results and commented on the manuscript. The authors declare that they have no conflict of interest.Attached Files
Published - embr.201847182.pdf
Submitted - 423905.full.pdf
Supplemental Material - embr201847182-sup-0001-appendix.pdf
Supplemental Material - embr201847182-sup-0002-movieev1.zip
Supplemental Material - embr201847182-sup-0003-movieev2.zip
Supplemental Material - embr201847182-sup-0004-movieev3.zip
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Additional details
- Eprint ID
- 90012
- Resolver ID
- CaltechAUTHORS:20180927-114225130
- ANR-14-CE19-0006-01
- Agence Nationale pour la Recherche (ANR)
- 772103-BRIDGING
- European Research Council (ERC)
- European Molecular Biology Organization (EMBO)
- NEAMEMB PDR T.1003.14
- Fonds National de la Recherche Scientifique (FNRS)
- BRIDGING CDR J.0114.18
- Fonds National de la Recherche Scientifique (FNRS)
- RHAMEMB CDR J.0086.18
- Fonds National de la Recherche Scientifique (FNRS)
- 1.E.096.18
- Formation à la Recherche dans l'Industrie et l'Agriculture
- BB/M007200/1
- Biotechnology and Biological Sciences Research Council (BBSRC)
- Scottish Government Rural and Environment Science and Analytical Services Division (RESAS)
- Created
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2018-09-27Created from EPrint's datestamp field
- Updated
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2023-06-01Created from EPrint's last_modified field