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Prohibitin is required for Ras-induced Raf–MEK–ERK activation and epithelial cell migration

Nature Cell Biology volume 7, pages 837–843 (2005)Cite this article

Abstract

Ras proteins control the signalling pathways that are responsible for normal growth and malignant transformation1. Raf protein kinases are direct Ras effector proteins that initiate the mitogen-activated protein kinase (MAPK) cascade2, which mediates diverse biological functions such as cell growth, survival and differentiation3. Here we show that prohibitin, a ubiquitously expressed and evolutionarily conserved protein4 is indispensable for the activation of the Raf–MEK–ERK pathway by Ras. The membrane targeting and activation of C-Raf by Ras needs prohibitin in vivo. In addition, direct interaction with prohibitin is required for C-Raf activation. C-Raf kinase fails to interact with the active Ras induced by epidermal growth factor in the absence of prohibitin. Moreover, in prohibitin-deficient cells the adhesion complex proteins cadherin and β-catenin relocalize to the plasma membrane and thereby stabilize adherens junctions. Our data show an unexpected role of prohibitin in the activation of the Ras–Raf signalling pathway and in modulating epithelial cell adhesion and migration.

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Figure 1: Knockdown of PHB induces change in epithelial cell morphology.
Figure 2: Knockdown of PHB induces adherens junction formation and changes in epithelial morphogenesis.
Figure 3: PHB is required for EGF-induced Raf–MAPK activation.
Figure 4: PHB is required for the activation of C-Raf by Ras.
Figure 5: PHB is required for 14-3-3 displacement from C-Raf.

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Acknowledgements

We thank M. Oswald, D. Khalil, C. Dimmler, B. Fauler and U. Reichard for excellent technical assistance, T. Fowler for critical reading of the manuscript and the EURIT team for their help with siRNA validation. M. Selbach is thanked for kindly providing the Ras constructs and S. Lohmann for the VASP-P157 antibodies. This work was supported by grants from the Bundesministerium für Bildung und Forschung (BMBF) to T.R.

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Authors and Affiliations

  1. Department of Molecular Biology, Max Planck Institute for Infection Biology, Schumannstr. 21/22, Berlin, D-10117, Germany

    Krishnaraj Rajalingam, Christian Wunder, Yuri Churin, Claudia Sievers & Thomas Rudel

  2. Microscopy Core Facility, Schumannstr. 21/22, Berlin, D-10117, Germany

    Volker Brinkmann

  3. Institut für Medizinische Strahlenkunde und Zellforschung, University of Würzburg, Versbacher Strasse 5, Würzburg, 97078, Germany

    Mirko Hekman & Ulf R. Rapp

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Correspondence to Thomas Rudel.

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Rajalingam, K., Wunder, C., Brinkmann, V. et al. Prohibitin is required for Ras-induced Raf–MEK–ERK activation and epithelial cell migration. Nat Cell Biol 7, 837–843 (2005). https://doi.org/10.1038/ncb1283

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