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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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.
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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