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Eps8 controls actin-based motility by capping the barbed ends of actin filaments

Nature Cell Biology volume 6, pages 1180–1188 (2004)Cite this article

Abstract

Actin filament barbed-end capping proteins are essential for cell motility, as they regulate the growth of actin filaments to generate propulsive force. One family of capping proteins, whose prototype is gelsolin, shares modular architecture, mechanism of action, and regulation through signalling-dependent mechanisms, such as Ca2+ or phosphatidylinositol-4,5-phosphate binding. Here we show that proteins of another family, the Eps8 family, also show barbed-end capping activity, which resides in their conserved carboxy-terminal effector domain. The isolated effector domain of Eps8 caps barbed ends with an affinity in the nanomolar range. Conversely, full-length Eps8 is auto-inhibited in vitro, and interaction with the Abi1 protein relieves this inhibition. In vivo, Eps8 is recruited to actin dynamic sites, and its removal impairs actin-based propulsion. Eps8-family proteins do not show any similarity to gelsolin-like proteins. Thus, our results identify a new family of actin cappers, and unveil novel modalities of regulation of capping through protein–protein interactions. One established function of the Eps8–Abi1 complex is to participate in the activation of the small GTPase Rac, suggesting a multifaceted role for this complex in actin dynamics, possibly through the participation in alternative larger complexes.

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Figure 1: Eps8 'output' region binds F-actin and possesses filament barbed end-capping activity.
Figure 2: Eps8 family members function as barbed-end-capping proteins.
Figure 3: The filament barbed-end capping activity of Eps8 permits the reconstitution of actin-based motility in vitro.
Figure 4: Eps8 is required for optimal actin-based motility in vivo.
Figure 5: Eps8 activity modulated by association with Abi1.

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Acknowledgements

This work was supported by grants: from AIRC (Associazione Italiana Ricerca sul Cancro) to G.S. and P.P.D.F.; from Human Science Frontier Program to G.S. and M.F.C. (grant RGP0072/2003-C), and to P.P.D.F.; from the Italian Ministry of Health (grant R.F. 02/184) to G.S.; from the 'French Ligue Nationale Contre le Cancer to M.F.C. 'équipe labellisée Ligue'; from the German research council (DFG, Str 666/1-1) to T.E.B.S.; from the Deutsche Akademische Austauschdienst (DAAD) and from the Fonds der Chemischen Industrie to J.W.; from European Community (VI Framework) to G.S. and P.P.D.F.; and from Fondazione Monzino to PPDF. We thank L. Cairns, P. Hagendorff and S. Bossi for technical help; and E. Helfer and S. Wiesner for assistance on actin polymerization and motility assays. We are grateful to T. Chakraborti for providing GFP-expressing L. monocytogenes and L. Machesky for Myc–PI(4,5)K. T.E.B.S. and D.D. contributed equally to this work.

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Author notes

  1. Andrea Disanza and Marie-France Carlier: These authors contributed equally to this work.

Authors and Affiliations

  1. IFOM Istituto FIRC di Oncologia Molecolare Via Adamello 16, Milan, 20139, Italy

    Andrea Disanza, Emanuela Frittoli, Stefano Confalonieri, Assunta Croce, Pier Paolo Di Fiore & Giorgio Scita

  2. Department of Experimental Oncology, Istituto Europeo di Oncologia (IEO), Via Ripamonti 435, Milan, 20141, Italy

    Andrea Disanza, Emanuela Frittoli, Stefano Confalonieri, Assunta Croce, Pier Paolo Di Fiore & Giorgio Scita

  3. Dynamique du Cytosquelette Laboratoire d'Enzymologie et Biochimie Structurales, C.N.R.S., Gif-sur -Yvette, 91198, France

    Andrea Disanza, Marie-France Carlier & Dominique Didry

  4. Department of Cell Biology, German Research Centre for Biotechnology (GBF), Mascheroder Weg 1, Braunschweig, D-38124, Germany

    Theresia E. B. Stradal & Jurgen Wehland

  5. Dipartimento di Medicina, Chirurgia ed Odontoiatria, Universita' degli Studi di Milano, Milan, 20122, Italy

    Pier Paolo Di Fiore

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Correspondence to Marie-France Carlier or Giorgio Scita.

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Disanza, A., Carlier, MF., Stradal, T. et al. Eps8 controls actin-based motility by capping the barbed ends of actin filaments. Nat Cell Biol 6, 1180–1188 (2004). https://doi.org/10.1038/ncb1199

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