Cell motility is essential for viral dissemination1. Vaccinia virus (VACV), a close relative of smallpox virus, is thought to exploit cell motility as a means to enhance the spread of infection1. A single viral protein, F11L, contributes to this by blocking RhoA signalling to facilitate cell retraction2. However, F11L alone is not sufficient for VACV-induced cell motility, indicating that additional viral factors must be involved. Here, we show that the VACV epidermal growth factor homologue, VGF, promotes infected cell motility and the spread of viral infection. We found that VGF secreted from early infected cells is cleaved by ADAM10, after which it acts largely in a paracrine manner to direct cell motility at the leading edge of infection. Real-time tracking of cells infected in the presence of EGFR, MAPK, FAK and ADAM10 inhibitors or with VGF-deleted and F11-deleted viruses revealed defects in radial velocity and directional migration efficiency, leading to impaired cell-to-cell spread of infection. Furthermore, intravital imaging showed that virus spread and lesion formation are attenuated in the absence of VGF. Our results demonstrate how poxviruses hijack epidermal growth factor receptor-induced cell motility to promote rapid and efficient spread of infection in vitro and in vivo.
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We thank K. Gonciarz and I. F. Sbalzarini for helpful discussions. We thank B. Siegenthaler and J. Crouse for initial work on the project. We thank all members of the Mercer lab for helpful discussions and comments on the manuscript. We thank A. Vaughan, V. Jäggin, T. Lopes, T. Lummen, T. Horn and D. Loeffler for technical support. We thank B. Moss (NIAID) for the vSC20 virus. This research was supported by core funding to the MRC Laboratory for Molecular Cell Biology, University College London (MC_UU12018/7) (J.M.), the European Research Council (649101–UbiProPox) (J.M.), ETH Zurich (G.F. and D.J.M.) and the Division of Intramural Research of the NIAID (G.V.R. and H.D.H.). C.B. is funded by the MRC Laboratory for Molecular Cell Biology PhD programme.
The authors declare no competing interests.
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Supplementary Figures 1–5, Supplementary Table 1.
VACV WR and mutant plaque formation.
VACV plaque formation in the presence of EGFR, MEK or FAK inhibitors.
VACV plaque formation adjacent to wounds.
VACV plaque formation in the presence of ADAM10 inhibitor.
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Beerli, C., Yakimovich, A., Kilcher, S. et al. Vaccinia virus hijacks EGFR signalling to enhance virus spread through rapid and directed infected cell motility. Nat Microbiol 4, 216–225 (2019). https://doi.org/10.1038/s41564-018-0288-2
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