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Synaptotagmin-like proteins control the formation of a single apical membrane domain in epithelial cells

Nature Cell Biology volume 14, pages 838–849 (2012)Cite this article

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Abstract

The formation of epithelial tissues requires both the generation of apical–basal polarity and the coordination of this polarity between neighbouring cells to form a central lumen. During de novo lumen formation, vectorial membrane transport contributes to the formation of a singular apical membrane, resulting in the contribution of each cell to only a single lumen. Here, from a functional screen for genes required for three-dimensional epithelial architecture, we identify key roles for synaptotagmin-like proteins 2-a and 4-a (Slp2-a/4-a) in the generation of a single apical surface per cell. Slp2-a localizes to the luminal membrane in a PtdIns(4,5)P2-dependent manner, where it targets Rab27-loaded vesicles to initiate a single lumen. Vesicle tethering and fusion is controlled by Slp4-a, in conjunction with Rab27/Rab3/Rab8 and the SNARE syntaxin-3. Together, Slp2-a/4-a coordinate the spatiotemporal organization of vectorial apical transport to ensure that only a single apical surface, and thus the formation of a single lumen, occurs per cell.

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Figure 1: A screen for regulators of 3D epithelial polarization.
Figure 2: Slp2-a is required for epithelial morphogenesis.
Figure 3: Slp2-a requires SHD and C2A/B domains for correct localization.
Figure 4: Slp2-a binds Rab27 to form the apical membrane.
Figure 5: Slp4-a is required for epithelial morphogenesis.
Figure 6: Slp4-a binding to the plasma membrane, Rabs and Stx3 is required for apical membrane formation.
Figure 7: Slp2-a regulates Slp4-a targeting to determine single apical membrane formation.

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Acknowledgements

We thank C. M. Ruiz-Jarabo for comments on the manuscript, and members of the Martin-Belmonte laboratory for discussion. We thank M. ter Beest, J. Peränen, and K. Simons, Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG), Dresden, Germany, for generous gifts of reagents, and the Mostov laboratory for kind assistance. This work was supported by grants from the Human Frontiers Science Program (HFSP-CDA 00011/2009), Marie Curie (IRG-209382), MICINN (BFU2008-01916), (BFU2011-22622) and CONSOLIDER (CSD2009-00016) to F.M-B.; by NIH R01DK074398, R01AI25144 and R01DK91530 to K.M., and The March of Dimes Basil O’Connor Starter Research Award to P.R.B. A.E.R-F. is the recipient of a JAE fellowship, from CSIC; M.G-S. is the recipient of a FPI fellowship, from MICINN; and I.B-R. is the recipient of an AECC fellowship. An institutional Grant from the Fundación Ramón Areces to CBMSO is also acknowledged.

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Author notes

  1. Silvia Vergarajauregui & Christiaan L. Slim

    Present address: Present addresses: National Heart, Lung and Blood Institute (NHLBI), National Institutes of Health (NIH) Bethesda, Maryland 20824-0105, USA (S.V.); Department of Cell Biology, University Medical Center Groningen, Antonius Deusinglaan 1, Groningen 9713 AV, Holland (C.L.S.),

  2. Manuel Gálvez-Santisteban and Alejo E. Rodriguez-Fraticelli: These authors contributed equally to this work

Authors and Affiliations

  1. Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas (CSIC), C/Nicolás Cabrera 1, Madrid 28049, Spain

    Manuel Gálvez-Santisteban, Alejo E. Rodriguez-Fraticelli, Silvia Vergarajauregui, Inmaculada Bañón-Rodríguez, Ilenia Bernascone & Fernando Martín-Belmonte

  2. Department of Anatomy, University of California San Francisco, California 94143-2140, USA

    David M. Bryant, Anirban Datta, Natalie Spivak, Kitty Young, Christiaan L. Slim, Paul R. Brakeman & Keith E. Mostov

  3. Department of Developmental Biology and Neurosciences, Graduate School of Life Sciences, Tohoku University, Aobayama, Aoba-ku, Sendai, Miyagi 980-8578, Japan

    Takao Yasuda & Mitsunori Fukuda

  4. Department of Pediatrics, University of California San Francisco, California 94143-2140, USA

    Natalie Spivak & Paul R. Brakeman

  5. Department of Biochemistry and Biophysics, University of California San Francisco, California 94143-2140, USA

    Keith E. Mostov

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  1. Manuel Gálvez-Santisteban
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Contributions

M.G-S., A.E.R-F., D.M.B., S.V. and F.M-B. designed the experiments. M.G-S., A.E.R-F., D.M.B., S.V., T.S., I.B.R., I.B., A.D., N.S., K.Y. and C.L.S. did the experimental work. M.G-S., A.E.R-F., D.M.B., K.E.M. and F.M-B. analysed the experiments. P.R.B. and M.F. provided reagents. F.M-B., D.M.B. M.G-S. and A.E.R-F. wrote the manuscript.

Corresponding author

Correspondence to Fernando Martín-Belmonte.

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The authors declare no competing financial interests.

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Gálvez-Santisteban, M., Rodriguez-Fraticelli, A., Bryant, D. et al. Synaptotagmin-like proteins control the formation of a single apical membrane domain in epithelial cells. Nat Cell Biol 14, 838–849 (2012). https://doi.org/10.1038/ncb2541

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