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Virus\u2010induced cell migration facilitates viral transmission. However, the mechanism underlying this type of cell migration remains unclear. Here we investigate the motility of cells infected by vaccinia virus (VACV), a close relative of monkeypox, through combining multi\u2010omics analyses and high\u2010resolution live\u2010cell imaging. We find that, upon VACV infection, the epithelial cells undergo epithelial\u2013mesenchymal transition\u2010like transformation, during which they lose intercellular junctions and acquire the migratory capacity to promote viral spreading. After transformation, VACV\u2010hijacked RhoA signaling significantly alters cellular morphology and rearranges the actin cytoskeleton involving the depolymerization of robust actin stress fibers, leading\u2010edge protrusion formation, and the rear\u2010edge recontraction, which coordinates VACV\u2010induced cell migration. Our study reveals how poxviruses alter the epithelial phenotype and regulate RhoA signaling to induce fast migration, providing a unique perspective to understand the pathogenesis of poxviruses.<\/jats:p>","DOI":"10.1002\/jmv.29041","type":"journal-article","created":{"date-parts":[[2023,8,25]],"date-time":"2023-08-25T02:42:35Z","timestamp":1692931355000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Vaccinia virus induces EMT\u2010like transformation and RhoA\u2010mediated mesenchymal migration"],"prefix":"10.1002","volume":"95","author":[{"given":"Wei","family":"Liu","sequence":"first","affiliation":[{"name":"Shanghai Xuhui Central Hospital, Zhongshan\u2010Xuhui Hospital, Shanghai Key Laboratory of Medical Epigenetics, Shanghai Stomatological Hospital, Institutes of Biomedical Sciences, Department of Chemistry Fudan University Shanghai 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