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RhoD regulates endosome dynamics through Diaphanous-related Formin and Src tyrosine kinase

Nature Cell Biology volume 5, pages 195–204 (2003)Cite this article

A Corrigendum to this article was published on 01 July 2003

Abstract

Early endosomes move bidirectionally between the cell periphery and the interior through a mechanism regulated by the low molecular weight GTPase RhoD. Here, we identify a novel splice variant of human Diaphanous, hDia2C, which specifically binds to RhoD and is recruited onto early endosomes. Expression of RhoD and hDia2C induces a striking alignment of early endosomes along actin filaments and reduces their motility. This activity depends on the membrane recruitment and activation of c-Src kinase, thus uncovering a new role in endosome function. Our results define a novel signal transduction pathway, in which hDia2C and c-Src are sequentially activated by RhoD to regulate the motility of early endosomes through interactions with the actin cytoskeleton.

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Figure 1: hDia2C, a new member of the Formin-Related family, is a RhoD effector.
Figure 2: The GTPase-deficient mutant RhoDG26V targets hDia2C on endosomes.
Figure 3: RhoD and hDia2C regulate endosomes motility in vivo.
Figure 4: RhoD and hDia2C regulate endosome–cytoskeleton interactions.
Figure 5: hDia2C stimulates Src-kinase activity in HeLa cells.
Figure 6: RhoD/hDia2C regulate endosomes motility through Src kinase.

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Acknowledgements

We are grateful to S. Narumiya for providing pFL-mDia1 and pFL-mDia1ΔN3, H. McBride for pLexA-RhoDG26V (−CAAX) and D. Toniolo for pCMV-HA-hDia2B. We acknowledge the generosity of P. Soriano and G. Superti-Furga for kindly providing Src+/+ and Src−/− cell lines, as well as pEGFP-SrcWT and pEGFP-SrcY527F. Special thanks A. Desay, J. Howard, H. Mellor, C. Murphy and J. Rink for valuable discussions and comments on the manuscript, and to S. Chasserot-Golaz for help with confocal microscopy. We would like also to thank A. Giner for technical assistance, B. Habermann for biocomputing analysis and D. Bagnard for sharing technical resources. This work was supported by a Human Frontier Science Program (HFSP) long-term fellowship to S.G. (LT0493/1999-M) and by a HFSP grant to M.Z. (RG0260/1999-M)

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Authors and Affiliations

  1. Max Planck Institute for Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, Dresden, D-01307, Germany

    Stéphane Gasman & Marino Zerial

  2. CNRS UPR-2356 Neurotransmission et Sécrétion Neuroendocrine, 5 rue Blaise Pascal, Strasbourg, 67084, France

    Stéphane Gasman

  3. A.N. Belozersky Institute of Physico-chemical Biology, Build A, Moscow State University, Moscow, 119899, Russia

    Yannis Kalaidzidis

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Correspondence to Marino Zerial.

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Supplementary information

Supplementary Movie

All videos were acquired and processed as described previously in Live Cell Imaging and Image Processing. They comprise 100 frames, animated at 20 frames/s (around 7 × acquisition time). (MOV 2390 kb)

Movie 1 HeLa cells co-overexpressing hRab5-GFP and empty pCDNA3.1.

Supplementary Movie

Movie 2 HeLa cells co-overexpressing hRab5-GFP and RhoDV26. (MOV 2326 kb)

Supplementary Movie

Movie 3 HeLa cells co-overexpressing hRab5-GFP and ΔGBD-hDia2C. (MOV 1898 kb)

Supplementary Figure

Figure S1 mDia1 do not regulate endosome-cytoskeleton interactions (PDF 176 kb)

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Gasman, S., Kalaidzidis, Y. & Zerial, M. RhoD regulates endosome dynamics through Diaphanous-related Formin and Src tyrosine kinase. Nat Cell Biol 5, 195–204 (2003). https://doi.org/10.1038/ncb935

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