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Phosphatidylinositol-3,4,5-trisphosphate regulates the formation of the basolateral plasma membrane in epithelial cells

Nature Cell Biology volume 8, pages 963–970 (2006)Cite this article

A Corrigendum to this article was published on 01 October 2006

Abstract

Polarity is a central feature of eukaryotic cells and phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P3) has a central role in the polarization of neurons and chemotaxing cells. In polarized epithelial cells, PtdIns(3,4,5)P3 is stably localized at the basolateral plasma membrane, but excluded from the apical plasma membrane, as shown by localization of GFP fused to the PtdIns(3,4,5)P3-binding pleckstrin-homology domain of Akt (GFP-PH–Akt), a fusion protein that indicates the location of PtdIns(3,4,5)P3. Here, we ectopically inserted exogenous PtdIns(3,4,5)P3 into the apical plasma membrane of polarized Madin-Darby canine kidney (MDCK) cells. Within 5 min many cells formed protrusions that extended above the apical surface. These protrusions contained basolateral plasma membrane proteins and excluded apical proteins, indicating that their plasma membrane was transformed from apical to basolateral. Addition of PtdIns(3,4,5)P3 to the basolateral surface of MDCK cells grown as cysts caused basolateral protrusions. MDCK cells grown in the presence of a phosphatidylinositol 3-kinase inhibitor had abnormally short lateral surfaces, indicating that PtdIns(3,4,5)P3 regulates the formation of the basolateral surface.

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Figure 1: Exogenous PtdIns(3,4,5)P3 delivered to the apical surface of MDCK cells induces protrusions.
Figure 2: Endogenous Rac1 and Cdc42 are activated by exogenous PtdIns(3,4,5)P3.
Figure 3: PtdIns(3,4,5)P3 regulates cell polarity.
Figure 4: PtdIns(3,4,5)P3 stimulates transcytosis.
Figure 5: PtdIns(3,4,5)P3 is required for formation and maintenance of the basolateral surface.

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Acknowledgements

We thank K. Matlin, G. Ojakian and B. Stevenson for reagents. We thank H. Bourne, O. Weiner, M. Zegers, P. Brakeman and members of our laboratory for advice and comments on the paper. and D. Mills for help with the manuscript. This work was supported by National Institutes of Health (NIH) grants to K.M. and J.E. W.Y. is supported by a fellowship from the National Kidney Foundation. F.M.-B. is supported by the Human Frontiers Science Program (HFSP).

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Author notes

  1. Martin ter Beest

    Present address: The Vontz Center, University of Cincinnati College of Medicine, 3125 Eden Avenue, ML0581, Cincinnati, OH 45267-0581, USA

Authors and Affiliations

  1. Department of Anatomy, University of California, San Francisco, 94143, CA, USA

    Ama Gassama-Diagne, Wei Yu, Martin ter Beest, Fernando Martin-Belmonte & Keith Mostov

  2. Institut Federatif de Recherche Claude de Preval, IFR30, INSERM Unite 563, Departement Lipoproteines et Mediateurs Lipidiques, Hopital Purpan, Toulouse Cedex, 31059, France

    Ama Gassama-Diagne

  3. Departments of Medicine, University of California, San Francisco, 94143, CA, USA

    Arlinet Kierbel & Joanne Engel

  4. Microbiology and Immunology, University of California, San Francisco, 94143, CA, USA

    Joanne Engel

  5. Biochemistry and Biophysics, University of California, San Francisco, 94143, CA, USA

    Keith Mostov

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Contributions

A.G. designed, performed and interpreted all of the experiments. W.Y. helped with the design of some experiments. M.t.B. and F.M.-B. made invaluable reagents. A.K. and J.E. helped with the interpretation of data and writing of the paper. K.M. conceived the experiments, helped with interpretation of data and wrote the paper.

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Correspondence to Ama Gassama-Diagne.

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Gassama-Diagne, A., Yu, W., ter Beest, M. et al. Phosphatidylinositol-3,4,5-trisphosphate regulates the formation of the basolateral plasma membrane in epithelial cells. Nat Cell Biol 8, 963–970 (2006). https://doi.org/10.1038/ncb1461

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