Abstract
Neural Wiskott–Aldrich syndrome protein (N-WASP) and WAVE are members of a family of proteins that use the Arp2/3 complex to stimulate actin assembly in actin-based motile processes. By entering into distinct macromolecular complexes, they act as convergent nodes of different signalling pathways. The role of WAVE in generating lamellipodial protrusion during cell migration is well established. Conversely, the precise cellular functions of N-WASP have remained elusive. Here, we report that Abi1, an essential component of the WAVE protein complex, also has a critical role in regulating N-WASP-dependent function. Consistently, Abi1 binds to N-WASP with nanomolar affinity and, cooperating with Cdc42, potently induces N-WASP activity in vitro. Molecular genetic approaches demonstrate that Abi1 and WAVE, but not N-WASP, are essential for Rac-dependent membrane protrusion and macropinocytosis. Conversely, Abi1 and N-WASP, but not WAVE, regulate actin-based vesicular transport, epidermal growth factor receptor (EGFR) endocytosis, and EGFR and transferrin receptor (TfR) cell-surface distribution. Thus, Abi1 is a dual regulator of WAVE and N-WASP activities in specific processes that are dependent on actin dynamics.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Pantaloni, D., Le Clainche, C. & Carlier, M. F. Mechanism of actin-based motility. Science 292, 1502–1506 (2001).
Pollard, T. D. & Borisy, G. G. Cellular motility driven by assembly and disassembly of actin filaments. Cell 112, 453–465 (2003).
Qualmann, B. & Kessels, M. M. Endocytosis and the cytoskeleton. Int. Rev. Cytol. 220, 93–144 (2002).
Takenawa, T. & Miki, H. WASP and WAVE family proteins: key molecules for rapid rearrangement of cortical actin filaments and cell movement. J. Cell Sci. 114, 1801–1809 (2001).
Moreau, V. et al. A complex of N-WASP and WIP integrates signalling cascades that lead to actin polymerization. Nature Cell Biol. 2, 441–448 (2000).
Stradal, T. E. et al. Regulation of actin dynamics by WASP and WAVE family proteins. Trends Cell Biol. 14, 303–311 (2004).
Kitamura, T. et al. Molecular cloning of p125Nap1, a protein that associates with an SH3 domain of Nck. Biochem. Biophys. Res. Commun. 219, 509–514 (1996).
Kobayashi, K. et al. p140Sra-1 (specifically Rac1-associated protein) is a novel specific target for Rac1 small GTPase. J. Biol. Chem. 273, 291–295 (1998).
Eden, S., Rohatgi, R., Podtelejnikov, A. V., Mann, M. & Kirschner, M. W. Mechanism of regulation of WAVE1-induced actin nucleation by Rac1 and Nck. Nature 418, 790–793 (2002).
Shi, Y., Alin, K. & Goff, S. P. Abl-interactor-1, a novel SH3 protein binding to the carboxy-terminal portion of the Abl protein, suppresses v-abl transforming activity. Genes Dev. 9, 2583–2597 (1995).
Innocenti, M. et al. Abi1 is essential for the formation and activation of a WAVE2 signalling complex. Nature Cell Biol. 6, 319–327 (2004).
Gautreau, A. et al. Purification and architecture of the ubiquitous Wave complex. Proc. Natl Acad. Sci. USA 101, 4379–4383 (2004).
Parsons, M. et al. Spatially distinct binding of Cdc42 to PAK1 and N-WASP in breast carcinoma cells. Mol. Cell Biol. 25, 1680–1695 (2005).
Suetsugu, S., Yamazaki, D., Kurisu, S. & Takenawa, T. Differential roles of WAVE1 and WAVE2 in dorsal and peripheral ruffle formation for fibroblast cell migration. Dev. Cell 5, 595–609 (2003).
Steffen, A. et al. Sra-1 and Nap1 link Rac to actin assembly driving lamellipodia formation. EMBO J. 23, 749–759 (2004).
Innocenti, M. et al. Phosphoinositide 3-kinase activates Rac by entering in a complex with Eps8, Abi1, and Sos-1. J. Cell Biol. 160, 17–23 (2003).
Disanza, A. et al. Eps8 controls actin-based motility by capping the barbed ends of actin filaments. Nature Cell Biol. 6, 1180–1188 (2004).
Lorenz, M., Yamaguchi, H., Wang, Y., Singer, R. H. & Condeelis, J. Imaging sites of N-Wasp activity in lamellipodia and invadopodia of carcinoma cells. Curr. Biol. 14, 697–703 (2004).
Bogdan, S. & Klambt, C. Kette regulates actin dynamics and genetically interacts with Wave and Wasp. Development 130, 4427–4437 (2003).
Carlier, M. F. et al. GRB2 links signaling to actin assembly by enhancing interaction of neural Wiskott-Aldrich syndrome protein (N-WASp) with actin-related protein (ARP2/3) complex. J. Biol. Chem. 275, 21946–21952 (2000).
Innocenti, M. et al. Mechanisms through which Sos-1 coordinates the activation of Ras and Rac. J. Cell Biol. 156, 125–136 (2002).
Fukuoka, M. et al. A novel neural Wiskott-Aldrich syndrome protein (N-WASP) binding protein, WISH, induces Arp2/3 complex activation independent of Cdc42. J. Cell Biol. 152, 471–482 (2001).
Snapper, S. B. et al. N-WASP deficiency reveals distinct pathways for cell surface projections and microbial actin-based motility. Nature Cell Biol. 3, 897–904 (2001).
Lommel, S. et al. Actin pedestal formation by enteropathogenic Escherichia coli and intracellular motility of Shigella flexneri are abolished in N-WASP-defective cells. EMBO Rep. 2, 850–857 (2001).
Taunton, J. Actin filament nucleation by endosomes, lysosomes and secretory vesicles. Curr. Opin. Cell Biol. 13, 85–91 (2001).
Rozelle, A. L. et al. Phosphatidylinositol 4,5-bisphosphate induces actin-based movement of raft-enriched vesicles through WASP-Arp2/3. Curr. Biol. 10, 311–320 (2000).
Bogdan, S., Stephan, R., Löbke, C., Mertens, A. & Klämbt, C. Abi activates WASP to promote sensory organ development. Nature Cell Biol. 7, 10.1038/ncb1305 (2005).
Kessels, M. M. & Qualmann, B. Syndapins integrate N-WASP in receptor-mediated endocytosis. EMBO J. 21, 6083–6094 (2002).
Naqvi, S. N., Zahn, R., Mitchell, D. A., Stevenson, B. J. & Munn, A. L. The WASp homologue Las17p functions with the WIP homologue End5p/verprolin and is essential for endocytosis in yeast. Curr. Biol. 8, 959–962 (1998).
Conner, S. D. & Schmid, S. L. Regulated portals of entry into the cell. Nature 422, 37–44 (2003).
Merrifield, C. J., Feldman, M. E., Wan, L. & Almers, W. Imaging actin and dynamin recruitment during invagination of single clathrin-coated pits. Nature Cell Biol. 4, 691–698 (2002).
Merrifield, C. J., Qualmann, B., Kessels, M. M. & Almers, W. Neural Wiskott Aldrich Syndrome Protein (N-WASP) and the Arp2/3 complex are recruited to sites of clathrin-mediated endocytosis in cultured fibroblasts. Eur. J. Cell Biol. 83, 13–18 (2004).
Benesch, S. et al. N-WASP deficiency impairs EGF internalization and actin assembly at clathrin-coated pits. J. Cell Sci. 118, 3103–3115 (2005).
Yarar, D., Waterman-Storer, C. M. & Schmid, S. L. A dynamic actin cytoskeleton functions at multiple stages of clathrin-mediated endocytosis. Mol. Biol. Cell 16, 964–975 (2005).
Merrifield, C. J., Perrais, D. & Zenisek, D. Coupling between clathrin-coated-pit invagination, cortactin recruitment, and membrane scission observed in live cells. Cell 121, 593–606 (2005).
Wiley, H. S. Trafficking of the ErbB receptors and its influence on signaling. Exp. Cell Res. 284, 78–88 (2003).
Suetsugu, S., Miki, H. & Takenawa, T. Identification of two human WAVE/SCAR homologues as general actin regulatory molecules which associate with the Arp2/3 complex. Biochem. Biophys. Res. Commun. 260, 296–302 (1999).
Kunda, P., Craig, G., Dominguez, V. & Baum, B. Abi, Sra1, and Kette control the stability and localization of SCAR/WAVE to regulate the formation of actin-based protrusions. Curr. Biol. 13, 1867–1875 (2003).
Pantaloni, D., Boujemaa, R., Didry, D., Gounon, P. & Carlier, M. F. The Arp2/3 complex branches filament barbed ends: functional antagonism with capping proteins. Nature Cell Biol. 2, 385–391 (2000).
Loisel, T. P., Boujemaa, R., Pantaloni, D. & Carlier, M. F. Reconstitution of actin-based motility of Listeria and Shigella using pure proteins. Nature 401, 613–616 (1999).
Egile, C. et al. Activation of the CDC42 effector N-WASP by the Shigella flexneri IcsA protein promotes actin nucleation by Arp2/3 complex and bacterial actin-based motility. J. Cell Biol. 146, 1319–1332 (1999).
Rubinson, D. A. et al. A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference. Nature Genet. 33, 401–406 (2003).
Haglund, K. et al. Multiple monoubiquitination of RTKs is sufficient for their endocytosis and degradation. Nature Cell Biol. 5, 461–466 (2003).
Acknowledgements
This work was supported by grants from: Associazione Italiana Ricerca sul Cancro (AIRC) and AIRC Regionale Lombardia to G.S.; Human Science Frontier Program to G.S. and M.F.C. (grant number RGP0072/2003-C); the Italian Ministry of Health (grant R.F. 02/184) to G.S.; the French Ligue Nationale Contre le Cancer to M.F.C. ('équipe labellisée Ligue'); the Fondazione Italiana Ricerca sul Cancro (FIRC) to M.I.; European Community (VI Framework) to G.S.; EMBO to M.H.; and from the Deutsche Forschungsgemeinschaft (DFG) (SPP 1150) to K.R. and T.E.B.S. We would like to thank M. Garre and S. Bossi for technical help, and S. Lommel for providing N-WASP constructs.
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Information
Supplementary figures S1a, S2a, S3a ,S4a and movie legends plus Supplementary Methods (PDF 470 kb)
Rights and permissions
About this article
Cite this article
Innocenti, M., Gerboth, S., Rottner, K. et al. Abi1 regulates the activity of N-WASP and WAVE in distinct actin-based processes. Nat Cell Biol 7, 969–976 (2005). https://doi.org/10.1038/ncb1304
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/ncb1304
This article is cited by
-
RBFOX2 deregulation promotes pancreatic cancer progression and metastasis through alternative splicing
Nature Communications (2023)
-
Orchestration of synaptic functions by WAVE regulatory complex-mediated actin reorganization
Experimental & Molecular Medicine (2023)
-
Src family kinases, adaptor proteins and the actin cytoskeleton in epithelial-to-mesenchymal transition
Cell Communication and Signaling (2021)
-
The roles and prognostic significance of ABI1-TSV-11 expression in patients with left-sided colorectal cancer
Scientific Reports (2021)
-
CRISPR/CAS9-mediated knockout of Abi1 inhibits p185Bcr-Abl-induced leukemogenesis and signal transduction to ERK and PI3K/Akt pathways
Journal of Hematology & Oncology (2020)