Studies of the actin-based motility of the intracellular pathogens Listeria monocytogenes and Shigella flexneri have provided important insight into the events occurring at the leading edges of motile cells1,2,3,4,5. Like the bacteria Listeria and Shigella, vaccinia virus, a relative of the causative agent of smallpox, uses actin-based motility to spread between cells6. In contrast to Listeria or Shigella, the actin-based motility of vaccinia is dependent on an unknown phosphotyrosine protein, but the underlying mechanism remains obscure7. Here we show that phosphorylation of tyrosine 112 in the viral protein A36R by Src-family kinases is essential for the actin-based motility of vaccinia. Tyrosine phosphorylation of A36R results in a direct interaction with the adaptor protein Nck8 and the recruitment of the Ena/VASP family member N-WASP9 to the site of actin assembly. We also show that Nck and N-WASP are essential for the actin-based motility of vaccinia virus. We suggest that vaccinia virus spreads by mimicking the signalling pathways that are normally involved in actin polymerization at the plasma membrane.
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We are grateful to H. Miki for N-WASP cDNA and N-WASP antibody. We thank G. Smith and R. Blasco for providing recombinant vaccinia strains; J. White, C. Blaumüller, A. Desai and A. Ploubidou for reading the manuscript; T. Harder for encouraging us to try PP1; and S. Guth for the ‘round the world’ PCR method. S.G. was supported by a fellowship from the Boncompagni-Ludovisi Foundation.
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Frischknecht, F., Moreau, V., Röttger, S. et al. Actin-based motility of vaccinia virus mimics receptor tyrosine kinase signalling. Nature 401, 926–929 (1999). https://doi.org/10.1038/44860
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