A complex of N-WASP and WIP integrates signalling cascades that lead to actin polymerization

Abstract

Wiskott–Aldrich syndrome protein (WASP) and N-WASP have emerged as key proteins connecting signalling cascades to actin polymerization. Here we show that the amino-terminal WH1 domain, and not the polyproline-rich region, of N-WASP is responsible for its recruitment to sites of actin polymerization during Cdc42-independent, actin-based motility of vaccinia virus. Recruitment of N-WASP to vaccinia is mediated by WASP-interacting protein (WIP), whereas in Shigella WIP is recruited by N-WASP. Our observations show that vaccinia and Shigella activate the Arp2/3 complex to achieve actin-based motility, by mimicking either the SH2/SH3-containing adaptor or Cdc42 signalling pathways to recruit the N-WASP–WIP complex. We propose that the N-WASP–WIP complex has a pivotal function in integrating signalling cascades that lead to actin polymerization.

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Figure 1: The WH1 domain mediates recruitement of N-WASP to vaccinia.
Figure 2: GFP–N-WASP can bind to Nck and Cdc42.
Figure 3: Actin-based motility of vaccinia is Cdc42-independent.
Figure 4: Mutations analogous to those in Wiskott–Aldrich syndrome abolish recruitment of the N-WASP WH1 domain.
Figure 5: WIP is required for vaccinia actin-tail formation.
Figure 6: Overexpression of WIP(WBD) inhibits vaccinia recruitment of N-WASP and blocks actin-tail formation.
Figure 7: WIP and Nck are recruited to vaccinia in the absence of N-WASP.
Figure 8: Shigella recruits WIP and Nck.

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Acknowledgements

We thank H. Miki (University of Tokyo) for providing rat N-WASP cDNA and anti-N-WASP antibody, and A. Hall (University College London) for human Cdc42 and Rac1 templates used for PCR and the GST–Cdc42(L61) expression construct. We also thank P. Sansonetti (Institut Pasteur, Paris), G. Smith (University of Oxford) K. Aktories (Freiburg, Germany), N. Le Bot (EMBL) and G. Stier (EMBL) for providing Shigella, vaccinia strain ΔA36R, Toxin B, anti-GFP antibody and modified pET24b vector, respectively. We thank A. Desai and G. Superti-Furga for critical reading of the manuscript.

Correspondence and requests for materials should be addressed to M.W.

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Correspondence to Michael Way.

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Moreau, V., Frischknecht, F., Reckmann, I. et al. A complex of N-WASP and WIP integrates signalling cascades that lead to actin polymerization. Nat Cell Biol 2, 441–448 (2000). https://doi.org/10.1038/35017080

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