Vaccinia virus, a close relative of the causative agent of smallpox, exploits actin polymerization to enhance its cell-to-cell spread. We show that actin-based motility of vaccinia is initiated only at the plasma membrane and remains associated with it. There must therefore be another form of cytoplasmic viral transport, from the cell centre, where the virus replicates, to the periphery. Video analysis reveals that GFP-labelled intracellular enveloped virus particles (IEVs) move from their perinuclear site of assembly to the plasma membrane on microtubules. We show that the viral membrane protein A36R, which is essential for actin-based motility of vaccinia, is also involved in microtubule-mediated movement of IEVs. We further show that conventional kinesin is recruited to IEVs via the light chain TPR repeats and is required for microtubule-based motility of the virus. Vaccinia thus sequentially exploits the microtubule and actin cytoskeletons to enhance its cell-to-cell spread.
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We thank L. Goldstein, G. B. Stokin and A. Kamal (UCSD, San Diego, USA) for antibodies against the heavy and light chains of conventional kinesin, as well as for the GST–KLC2 expression construct. We also thank A. Matus (Friedrich Miescher Institute, Basel, Switzerland) for providing his unpublished pBactin-mb5tubulin-EGFP expression vector and A. Desai (Max Planck Institute for Molecular Cell Biology and Genetics, Dresden, Germany) for the HIPYER antibody. We also thank I. Vernos, D. Brunner, F. Valderrama, E. Piddini and N. Scaplehorn for suggestions and comments concerning the manuscript. A. P. is supported by a European Commission Marie Curie Individual fellowship. A. H. is supported by Wenner-Gren Foundation (Sweden).
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Rietdorf, J., Ploubidou, A., Reckmann, I. et al. Kinesin-dependent movement on microtubules precedes actin-based motility of vaccinia virus. Nat Cell Biol 3, 992–1000 (2001). https://doi.org/10.1038/ncb1101-992
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