{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,19]],"date-time":"2026-05-19T06:02:50Z","timestamp":1779170570731,"version":"3.51.4"},"reference-count":43,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2018,3,5]],"date-time":"2018-03-05T00:00:00Z","timestamp":1520208000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000925","name":"NHMRC","doi-asserted-by":"publisher","award":["APP100790"],"award-info":[{"award-number":["APP100790"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000925","name":"NHMRC","doi-asserted-by":"publisher","award":["GNT0632785"],"award-info":[{"award-number":["GNT0632785"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Ectromelia virus (ECTV) is an orthopoxvirus and the causative agent of mousepox. Like other poxviruses such as variola virus (agent of smallpox), monkeypox virus and vaccinia virus (the live vaccine for smallpox), ECTV promotes actin-nucleation at the surface of infected cells during virus release. Homologs of the viral protein A36 mediate this function through phosphorylation of one or two tyrosine residues that ultimately recruit the cellular Arp2\/3 actin-nucleating complex. A36 also functions in the intracellular trafficking of virus mediated by kinesin-1. Here, we describe the generation of a recombinant ECTV that is specifically disrupted in actin-based motility allowing us to examine the role of this transport step in vivo for the first time. We show that actin-based motility has a critical role in promoting the release of virus from infected cells in vitro but plays a minor role in virus spread in vivo. It is likely that loss of microtubule-dependent transport is a major factor for the attenuation observed when A36R is deleted.<\/jats:p>","DOI":"10.3390\/v10030111","type":"journal-article","created":{"date-parts":[[2018,3,6]],"date-time":"2018-03-06T07:37:25Z","timestamp":1520321845000},"page":"111","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Loss of Actin-Based Motility Impairs Ectromelia Virus Release In Vitro but Is Not Critical to Spread In Vivo"],"prefix":"10.3390","volume":"10","author":[{"given":"Melanie","family":"Duncan","sequence":"first","affiliation":[{"name":"School of Life and Environmental Sciences, The University of Sydney, Sydney, NSW 2006, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jacquelyn","family":"Horsington","sequence":"additional","affiliation":[{"name":"School of Life and Environmental Sciences, The University of Sydney, Sydney, NSW 2006, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Preethi","family":"Eldi","sequence":"additional","affiliation":[{"name":"School of Medicine, College of Health and Medicine, The University of Tasmania, Hobart, TAS 7005, Australia"},{"name":"The John Curtin School of Medical Research, Australian National University, Canberra, ACT 2601, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zahrah","family":"Al Rumaih","sequence":"additional","affiliation":[{"name":"School of Medicine, College of Health and Medicine, The University of Tasmania, Hobart, TAS 7005, Australia"},{"name":"The John Curtin School of Medical Research, Australian National University, Canberra, ACT 2601, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gunasegaran","family":"Karupiah","sequence":"additional","affiliation":[{"name":"School of Medicine, College of Health and Medicine, The University of Tasmania, Hobart, TAS 7005, Australia"},{"name":"The John Curtin School of Medical Research, Australian National University, Canberra, ACT 2601, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2193-596X","authenticated-orcid":false,"given":"Timothy","family":"Newsome","sequence":"additional","affiliation":[{"name":"School of Life and Environmental Sciences, The University of Sydney, Sydney, NSW 2006, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2018,3,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2645","DOI":"10.1099\/vir.0.81090-0","article-title":"Ectromelia virus: The causative agent of mousepox","volume":"86","author":"Esteban","year":"2005","journal-title":"J. Gen. Virol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"7427","DOI":"10.1128\/JVI.06636-11","article-title":"Loss of cytoskeletal transport during egress critically attenuates ectromelia virus infection in vivo","volume":"86","author":"Lynn","year":"2012","journal-title":"J. Virol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/j.semcdb.2015.10.008","article-title":"Viruses that ride on the coat-tails of actin nucleation","volume":"46","author":"Newsome","year":"2015","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1016\/j.chom.2013.08.011","article-title":"Arp2\/3-mediated actin-based motility: A tail of pathogen abuse","volume":"14","author":"Welch","year":"2013","journal-title":"Cell Host Microbe"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1128\/JVI.01814-10","article-title":"Variola and Monkeypox Viruses Utilize Conserved Mechanisms of Virion Motility and Release That Depend on Abl and Src Family Tyrosine Kinases","volume":"85","author":"Reeves","year":"2011","journal-title":"J. Virol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2161","DOI":"10.1007\/s00705-006-0791-2","article-title":"Ultrastructural study of myxoma virus morphogenesis","volume":"151","author":"Duteyrat","year":"2006","journal-title":"Arch. Virol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1279","DOI":"10.1099\/vir.0.79863-0","article-title":"Yaba-like disease virus protein Y144R, a member of the complement control protein family, is present on enveloped virions that are associated with virus-induced actin tails","volume":"85","author":"Law","year":"2004","journal-title":"J. Gen. Virol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"472","DOI":"10.1016\/j.tim.2008.07.009","article-title":"Vaccinia virus morphogenesis and dissemination","volume":"16","author":"Roberts","year":"2008","journal-title":"Trends Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1128\/jvi.68.1.130-147.1994","article-title":"Assembly of vaccinia virus: The second wrapping cisterna is derived from the trans Golgi network","volume":"68","author":"Schmelz","year":"1994","journal-title":"J. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2915","DOI":"10.1099\/0022-1317-83-12-2915","article-title":"The formation and function of extracellular enveloped vaccinia virus","volume":"83","author":"Smith","year":"2002","journal-title":"J. Gen. Virol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2961","DOI":"10.1099\/vir.0.81260-0","article-title":"Vaccinia virus intracellular enveloped virions move to the cell periphery on microtubules in the absence of the A36R protein","volume":"86","author":"Roberts","year":"2005","journal-title":"J. Gen. Virol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"11651","DOI":"10.1128\/JVI.75.23.11651-11663.2001","article-title":"Vaccinia virus intracellular movement is associated with microtubules and independent of actin tails","volume":"75","author":"Ward","year":"2001","journal-title":"J. Virol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"4523","DOI":"10.1038\/emboj.2011.326","article-title":"A kisensin-1 binding motif in vaccinia virus that is widespread throughout the human genome","volume":"30","author":"Dodding","year":"2011","journal-title":"EMBO J."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1111\/tra.12494","article-title":"Vaccinia virus proteins A36 and F12\/E2 show strong preferences for different kinesin light chain isoforms","volume":"18","author":"Gao","year":"2017","journal-title":"Traffic"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Horsington, J., Lynn, H., Turnbull, L., Cheng, D., Braet, F., Diefenbach, R.J., Whitchurch, C.B., Karupiah, G., and Newsome, T.P. (2013). A36-dependent actin filament nucleation promotes release of vaccinia virus. PLoS Pathog., 9.","DOI":"10.1371\/journal.ppat.1003239"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1111\/j.1462-5822.2005.00613.x","article-title":"Abl collaborates with Src family kinases to stimulate actin-based motility of vaccinia virus","volume":"8","author":"Newsome","year":"2006","journal-title":"Cell Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"430","DOI":"10.1111\/j.1469-0691.2005.01146.x","article-title":"Undertsanding how Listeria monocytogenes targets and crosses host barriers","volume":"11","author":"Lecuit","year":"2005","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1038\/385265a0","article-title":"Actin polymerisation is induced by Arp2\/3 protein complex at the surface of Listeria monocytogenes","volume":"385","author":"Welch","year":"1997","journal-title":"Nature"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"873","DOI":"10.1126\/science.1183173","article-title":"Repulsion of superinfecting virions: A mechanism for rapid virus spread","volume":"327","author":"Doceul","year":"2010","journal-title":"Science"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1006\/viro.1994.1542","article-title":"Vaccinia virus gene A36R encodes a M, 43\u201350 K protein on the surface of extracellular enveloped virus","volume":"204","author":"Parkinson","year":"1994","journal-title":"Virology"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1038\/nature07773","article-title":"The rate of N-WASP exchange limits the extent of ARP2\/3-complex-dependent actin-based motility","volume":"458","author":"Weisswange","year":"2009","journal-title":"Nature"},{"key":"ref_22","first-page":"15","article-title":"Construction and Isolation of Recombinant Vaccinia Virus Using Genetic Markers","volume":"Volume 269","author":"Isaacs","year":"2004","journal-title":"Vaccinia Virus and Poxvirology Methods and Protocols"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"636","DOI":"10.1038\/378636a0","article-title":"Actin-based motilty of vaccinia virus","volume":"378","author":"Cudmore","year":"1995","journal-title":"Nature"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"16141","DOI":"10.1038\/nmicrobiol.2016.141","article-title":"NPF motifs in the vaccinia virus protein A36 recruit intersectin-1 to promote Cdc42:N-WASP-mediated viral release from infected cells","volume":"1","author":"Snetkov","year":"2016","journal-title":"Nat. Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"926","DOI":"10.1038\/44860","article-title":"Actin-based motility of vaccinia virus mimics receptor tyrosine kinase signalling","volume":"401","author":"Frischknecht","year":"1999","journal-title":"Nature"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1126\/science.1101509","article-title":"SRC mediates a switch from microtubule to actin-based motility of vaccinia virus","volume":"306","author":"Newsome","year":"2004","journal-title":"Science"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"41","DOI":"10.4149\/av_2010_01_41","article-title":"Contribution of rearranged actin structures to the spread of Ectromelia virus","volume":"54","author":"Boratynska","year":"2010","journal-title":"Acta Virol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"536","DOI":"10.1016\/j.chom.2009.10.011","article-title":"Nck- and N-WASP-dependent actin-based motility is conserved in divergent vertebrate poxviruses","volume":"6","author":"Dodding","year":"2009","journal-title":"Cell Host Microbe"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3389","DOI":"10.1093\/nar\/25.17.3389","article-title":"Gapped BLAST and PSI-BLAST: A new generation of protein database search programs","volume":"25","author":"Altschul","year":"1997","journal-title":"Nucleic Acids Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1006\/viro.2000.0761","article-title":"The genome sequence of Yaba-like disease virus, a yatapoxvirus","volume":"281","author":"Lee","year":"2001","journal-title":"Virology"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"731","DOI":"10.1038\/nm1265","article-title":"Disabling poxvirus pathogenesis by inhibition of Abl-family tyrosine kinases","volume":"11","author":"Reeves","year":"2005","journal-title":"Nat. Med."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1876","DOI":"10.1099\/vir.0.043943-0","article-title":"Protein B5 is required on extracellular enveloped vaccinia virus for repulsion of superinfecting virions","volume":"93","author":"Doceul","year":"2012","journal-title":"J. Gen. Virol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"3319","DOI":"10.1128\/jvi.67.6.3319-3325.1993","article-title":"Dissociation of progeny vaccinia virus from the cell-membrane is regulated by a viral envelope glycoprotien\u2014Effect of a point mutation in the lectin homology domain of the A34R gene","volume":"67","author":"Blasco","year":"1993","journal-title":"J. Virol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1016\/j.antiviral.2005.11.006","article-title":"Pathogenicity and immunogenicity in mice of vaccinia viruses mutated in the viral envelope proteins A33R and B5R","volume":"69","author":"Gurt","year":"2006","journal-title":"Antivir. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"12266","DOI":"10.1128\/JVI.77.22.12266-12275.2003","article-title":"Mutations in the vaccinia virus A33R and B5R envelope proteins that enhance release of extracellular virions and eliminate formation of actin-containing microvilli without preventing tyrosine phosphorylation of the A36R protein","volume":"77","author":"Katz","year":"2003","journal-title":"J. Virol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"11637","DOI":"10.1128\/JVI.76.22.11637-11644.2002","article-title":"Identification of Second-Site Mutations That Enhance Release and Spread of Vaccinia Virus","volume":"76","author":"Katz","year":"2002","journal-title":"J. Virol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2150","DOI":"10.1128\/JVI.01969-07","article-title":"Vaccinia virus A34 glycoprotein determines the protein composition of the extracellular virus envelope","volume":"82","author":"Perdiguero","year":"2008","journal-title":"J. Virol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1099\/0022-1317-50-1-89","article-title":"Significance of extracellular enveloped virus in the in vitro and in vivo dissemination of Vaccinia","volume":"50","author":"Payne","year":"1980","journal-title":"J. Gen. Virol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"792","DOI":"10.1099\/vir.0.008169-0","article-title":"Smallpox virus plaque phenotypes: Genetic, geographical and case fatality relationships","volume":"90","author":"Olson","year":"2009","journal-title":"J. Gen. Virol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"643","DOI":"10.1099\/0022-1317-66-3-643","article-title":"Extracellular release of enveloped vaccinia virus from mouse nasal epithelial cells in vivo","volume":"66","author":"Payne","year":"1985","journal-title":"J. Gen. Virol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"288","DOI":"10.1002\/jcp.20721","article-title":"Actin-based motility of intracellular bacteria, and polarized surface distribution of the bacterial effector molecules","volume":"209","author":"Carlsson","year":"2006","journal-title":"J. Cell. Physiol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1038\/nrmicro1320","article-title":"Actin-dependent movement of bacterial pathogens","volume":"4","author":"Stevens","year":"2006","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Travier, L., Guadagnini, S., Gouin, E., Dufour, A., Chenal-Francisque, V., Cossart, P., Olivo-Marin, J.-C., Ghigo, J.-M., Disson, O., and Lecuit, M. (2013). ActA promotes Listera monocytogenes aggregation, intestinal colonization and carriage. PLoS Pathog., 9.","DOI":"10.1371\/journal.ppat.1003131"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/10\/3\/111\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T14:57:35Z","timestamp":1760194655000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/10\/3\/111"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,3,5]]},"references-count":43,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2018,3]]}},"alternative-id":["v10030111"],"URL":"https:\/\/doi.org\/10.3390\/v10030111","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,3,5]]}}}