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N-WASP deficiency reveals distinct pathways for cell surface projections and microbial actin-based motility

Nature Cell Biology volume 3, pages 897–904 (2001)Cite this article

Abstract

The Wiskott–Aldrich syndrome protein (WASP) family of molecules integrates upstream signalling events with changes in the actin cytoskeleton. N-WASP has been implicated both in the formation of cell-surface projections (filopodia) required for cell movement and in the actin-based motility of intracellular pathogens. To examine N-WASP function we have used homologous recombination to inactivate the gene encoding murine N-WASP. Whereas N-WASP-deficient embryos survive beyond gastrulation and initiate organogenesis, they have marked developmental delay and die before embryonic day 12. N-WASP is not required for the actin-based movement of the intracellular pathogen Listeria but is absolutely required for the motility of Shigella and vaccinia virus. Despite these distinct defects in bacterial and viral motility, N-WASP-deficient fibroblasts spread by using lamellipodia and can protrude filopodia. These results imply a crucial and non-redundant role for N-WASP in murine embryogenesis and in the actin-based motility of certain pathogens but not in the general formation of actin-containing structures.

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Figure 1: Targeted disruption of N-WASP by homologous recombination.
Figure 2: N-WASP deficiency results in neural-tube defects and developmental delay.
Figure 3: The actin-based motility of Shigella flexneri and vaccinia, but not that of Listeria, requires N-WASP.
Figure 4: WASP compensates for N-WASP in the actin-based motility of vaccinia but not that of Shigella.
Figure 5: N-WASP−/− ES-FLCs form lamellipodia and filopodia in response to stimulation with serum and with PDGF.
Figure 6: Normal serum-induced and growth-factor-induced actin assembly and barbed-end nucleation in N-WASP-deficient SV-FLCs.

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Acknowledgements

We thank M. Way, H. Miki, T. Takenawa, R. Rohatgi, M. Kirshner and R. Xavier for reagents; Y. Fujiwara and S. Orkin for assistance with in situ hybridization experiments; Y. Ohta for microinjection expertise; M. Charles for assistance in Shigella infections; W. Swat and F. Chen for critical suggestions on the manuscript; and members of the Snapper, Alt and Rosen laboratories for continued support. This work was supported by grants from the National Institutes of Health (NIH) (to S.B.S., F.S.R., S.M.T. and F.W.A.), the Howard Hughes Medical Institute (to F.W.A.), the Massachusetts General Hospital NIH Center for the Study of Inflammatory Bowel Disease (to S.B.S.), and the 2nd Department of Internal Medicine, Nagasaki University School of Medicine (to F.T.).

Author information

Authors and Affiliations

  1. Center for Blood Research, 200 Longwood Avenue, Boston, 02115, Massachusetts, USA

    Scott B. Snapper, Ching-Hui Liu, Darryll Dudley, Hunter Fraser, Fred S. Rosen & Frederick W. Alt

  2. Gastrointestinal Unit (Medical Services) and the Center for the Study of Inflammatory Bowel Disease, Massachusetts General Hospital, 55 Fruit Street, Boston, 02114, Massachusetts, USA

    Scott B. Snapper, Fuminao Takeshima, Ching-Hui Liu & Deanna Nguyen

  3. Department of Medicine, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Scott B. Snapper, Fuminao Takeshima, Sheila M. Thomas & Marcia B. Goldberg

  4. Department of Pediatrics, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Inés Antón, Christoph Klein, Raif Geha & Fred S. Rosen

  5. Division of Immunology, Howard Hughes Medical Institute, Children's Hospital, Boston, 02115, Massachusetts, USA

    Inés Antón & Raif Geha

  6. Hematology Division, Beth Israel Deaconess Hospital, Boston, 02215, Massachusetts, USA

    Sheila M. Thomas

  7. Department of Cell Biology, Harvard Medical School, Boston, 02115, Massachusetts, USA

    John H. Hartwig

  8. Department of Infectious Disease, University of Florida College of Medicine, Gainesville, 32601, Florida, USA

    Daniel Purich & Fred Southwick

  9. Cardiology Division, Brigham & Women's Hospital, Longwood Avenue, Boston, 02115, Massachusetts, USA

    Marco Lopez-Ilasaca

  10. Division of Pediatric Hematology/Oncology, Howard Hughes Medical Institute, Children's Hospital, Boston, 02115, Massachusetts, USA

    Christoph Klein

  11. Howard Hughes Medical Institute, Children's Hospital, Boston, 02115, Massachusetts, USA

    Laurie Davidson, Richard C. Mulligan & Frederick W. Alt

  12. Department of Pathology, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Roderick Bronson

  13. Department of Genetics, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Richard C. Mulligan & Frederick W. Alt

  14. Infectious Disease Unit, Massachusetts General Hospital, 55 Fruit Street, Boston, 02114, Massachusetts, USA

    Marcia B. Goldberg

  15. Hematology Division, Brigham & Women's Hospital, Longwood Avenue, Boston, 02115, Massachusetts, USA

    John H. Hartwig

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Supplementary figures

Figure S1 Nck and WIP are recruited to sites of actin assembly in Shigella- and vaccinia-infected SV-FLCs. (PDF 175 kb)

Figure S2 Neither Nck nor WIP is recruited to Shigella and vaccinia in the absence of N-WASP.

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Snapper, S., Takeshima, F., Antón, I. et al. N-WASP deficiency reveals distinct pathways for cell surface projections and microbial actin-based motility. Nat Cell Biol 3, 897–904 (2001). https://doi.org/10.1038/ncb1001-897

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