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WIP regulates N-WASP-mediated actin polymerization and filopodium formation

Nature Cell Biology volume 3, pages 484–491 (2001)Cite this article

Abstract

Induction of filopodia is dependent on activation of the small GTPase Cdc42 and on neural Wiskott–Aldrich-syndrome protein (N-WASP). Here we show that WASP-interacting protein (WIP) interacts directly with N-WASP and actin. WIP retards N-WASP/Cdc42-activated actin polymerization mediated by the Arp2/3 complex, and stabilizes actin filaments. Microinjection of WIP into NIH 3T3 fibroblasts induces filopodia; this is inhibited by microinjection of anti-N-WASP antibody. Microinjection of anti-WIP antibody inhibits induction of filopodia by bradykinin, by an active Cdc42 mutant (Cdc42(V12)) and by N-WASP. Our results indicate that WIP and N-WASP may act as a functional unit in filopodium formation, which is consistent with their role in actin-tail formation in cells infected with vaccinia virus or Shigella.

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Figure 1: Interaction of WIP and N-WASP.
Figure 2: In vitro interaction of WIP, N-WASP and Cdc42.
Figure 3: Binding of WIP to G- and F-actin.
Figure 4: Effect of WIP on N-WASP-induced activation of the Arp2/3 complex.
Figure 5: Effect of WIP on disassembly of actin filaments.
Figure 6: WIP induces filopodium formation and localizes to filopodia after bradykinin stimulation.
Figure 7: WIP and N-WASP act as a unit in filopodium formation.
Figure 8: WIP is a component of the bradykinin pathway.

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Acknowledgements

We thank R. Manoukian and M. Byrne for technical help, A. Hall for help with microinjections, M. Kirschner and L. Ma for discussions and reagents, and K. Barkalow for help with video microscopy. This work was supported by grants from the NIH, the March of Dimes and the Baxter, Olsten and Centeon Corporations. N.M-Q. is supported by a MEC postdoctoral grant, I.M.A. is supported by a Lady Tata fellowship, and R.R. is a member of the Medical Scientist Training Program at Harvard Medical School.

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Authors and Affiliations

  1. Department of Pediatrics, Division of Immunology, Children's Hospital, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Narcisa Martinez-Quiles, Inés M. Antón, Stephen P. Saville, Raif S. Geha & Narayanaswamy Ramesh

  2. Department of Cell Biology, Boston, 02115, Massachusetts, USA

    Rajat Rohatgi

  3. Department of Neurology, Boston, 02115, Massachusetts, USA

    Miguel Medina

  4. Department of Biochemistry, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, 108, Tokyo, Japan

    Hiroaki Miki, Hideki Yamaguchi & Tadaomi Takenawa

  5. Department of Medicine, Division of Experimental Medicine, Brigham and Women's Hospital, Boston, 02115, Massachusetts, USA

    John H. Hartwig

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Correspondence to Raif S. Geha.

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Martinez-Quiles, N., Rohatgi, R., Antón, I. et al. WIP regulates N-WASP-mediated actin polymerization and filopodium formation. Nat Cell Biol 3, 484–491 (2001). https://doi.org/10.1038/35074551

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