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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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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

Affiliations

  1. Center for Blood Research, 200 Longwood Avenue, Boston, Massachusetts 02115, 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, Massachusetts, 02114, USA

    • Scott B. Snapper
    • , Fuminao Takeshima
    • , Ching-Hui Liu
    •  & Deanna Nguyen
  3. Department of Medicine, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Scott B. Snapper
    • , Fuminao Takeshima
    • , Sheila M. Thomas
    •  & Marcia B. Goldberg
  4. Department of Pediatrics, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Inés Antón
    • , Christoph Klein
    • , Raif Geha
    •  & Fred S. Rosen
  5. Division of Immunology, Howard Hughes Medical Institute, Children's Hospital, Boston, Massachusetts 02115, USA

    • Inés Antón
    •  & Raif Geha
  6. Hematology Division, Beth Israel Deaconess Hospital, Boston, Massachusetts 02215, USA

    • Sheila M. Thomas
  7. Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, 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, Massachusetts 02115, USA

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

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

    • Laurie Davidson
    • , Richard C. Mulligan
    •  & Frederick W. Alt
  12. Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115, USA

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

    • Richard C. Mulligan
    •  & Frederick W. Alt
  14. Infectious Disease Unit, Massachusetts General Hospital, 55 Fruit Street, Boston, Massachusetts, 02114, USA

    • Marcia B. Goldberg
  15. Hematology Division, Brigham & Women's Hospital, Longwood Avenue, Boston, Massachusetts 02115, 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.Figure S2 Neither Nck nor WIP is recruited to Shigella and vaccinia in the absence of N-WASP.

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DOI

https://doi.org/10.1038/ncb1001-897