{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T02:05:38Z","timestamp":1790388338972,"version":"4.1.0"},"reference-count":53,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2020,7,19]],"date-time":"2020-07-19T00:00:00Z","timestamp":1595116800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100009139","name":"Deutsches Zentrum f\u00fcr Infektionsforschung","doi-asserted-by":"publisher","award":["FKZ8009801908"],"award-info":[{"award-number":["FKZ8009801908"]}],"id":[{"id":"10.13039\/100009139","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>The intracellular transport of nucleocapsids of the highly pathogenic Marburg, as well as Ebola virus (MARV, EBOV), represents a critical step during the viral life cycle. Intriguingly, a population of these nucleocapsids is distributed over long distances in a directed and polar fashion. Recently, it has been demonstrated that the intracellular transport of filoviral nucleocapsids depends on actin polymerization. While it was shown that EBOV requires Arp2\/3-dependent actin dynamics, the details of how the virus exploits host actin signaling during intracellular transport are largely unknown. Here, we apply a minimalistic transfection system to follow the nucleocapsid-like structures (NCLS) in living cells, which can be used to robustly quantify NCLS transport in live cell imaging experiments. Furthermore, in cells co-expressing LifeAct, a marker for actin dynamics, NCLS transport is accompanied by pulsative actin tails appearing on the rear end of NCLS. These actin tails can also be preserved in fixed cells, and can be visualized via high resolution imaging using STORM in transfected, as well as EBOV infected, cells. The application of inhibitory drugs and siRNA depletion against actin regulators indicated that EBOV NCLS utilize the canonical Arp2\/3-Wave1-Rac1 pathway for long-distance transport in cells. These findings highlight the relevance of the regulation of actin polymerization during directed EBOV nucleocapsid transport in human cells.<\/jats:p>","DOI":"10.3390\/cells9071728","type":"journal-article","created":{"date-parts":[[2020,7,21]],"date-time":"2020-07-21T06:38:55Z","timestamp":1595313535000},"page":"1728","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["Ebola Virus Nucleocapsid-Like Structures Utilize Arp2\/3 Signaling for Intracellular Long-Distance Transport"],"prefix":"10.3390","volume":"9","author":[{"given":"Katharina","family":"Grikscheit","sequence":"first","affiliation":[{"name":"Institute of Virology, Philipps University Marburg, Hans-Meerwein-Str. 2, 35043 Marburg, Germany"},{"name":"German Center for Infection Research (DZIF), Partner Site: Giessen-Marburg-Langen, Hans-Meerwein-Str. 2, 35043 Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7739-4798","authenticated-orcid":false,"given":"Olga","family":"Dolnik","sequence":"additional","affiliation":[{"name":"Institute of Virology, Philipps University Marburg, Hans-Meerwein-Str. 2, 35043 Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuki","family":"Takamatsu","sequence":"additional","affiliation":[{"name":"Institute of Virology, Philipps University Marburg, Hans-Meerwein-Str. 2, 35043 Marburg, Germany"},{"name":"Department of Virology I, National Institute of Infectious Diseases, Tokyo 208-0011, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana Raquel","family":"Pereira","sequence":"additional","affiliation":[{"name":"Oxford Nano Imager, Linacre House, Banbury Rd, Oxford OX2 8TA, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephan","family":"Becker","sequence":"additional","affiliation":[{"name":"Institute of Virology, Philipps University Marburg, Hans-Meerwein-Str. 2, 35043 Marburg, Germany"},{"name":"German Center for Infection Research (DZIF), Partner Site: Giessen-Marburg-Langen, Hans-Meerwein-Str. 2, 35043 Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,7,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1038\/nrmicro2574","article-title":"Subversion of the actin cytoskeleton during viral infection","volume":"9","author":"Taylor","year":"2011","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_2","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_3","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1093\/infdis\/jiv083","article-title":"Transport of Ebolavirus Nucleocapsids Is Dependent on Actin Polymerization: Live-Cell Imaging Analysis of Ebolavirus-Infected Cells","volume":"212","author":"Schudt","year":"2015","journal-title":"J. Infect. Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"14402","DOI":"10.1073\/pnas.1307681110","article-title":"Live-cell imaging of Marburg virus-infected cells uncovers actin-dependent transport of nucleocapsids over long distances","volume":"113","author":"Schudt","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1075","DOI":"10.1073\/pnas.1712263115","article-title":"Ebola virus proteins NP, VP35, and VP24 are essential and sufficient to mediate nucleocapsid transport","volume":"115","author":"Takamatsu","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1146\/annurev-pathol-052016-100506","article-title":"The Pathogenesis of Ebola Virus Disease","volume":"12","author":"Baseler","year":"2017","journal-title":"Annu. Rev. Pathol."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Cantoni, D., and Rossman, J.S. (2018). Ebolaviruses: New roles for old proteins. PLoS Negl. Trop. Dis., 12.","DOI":"10.1371\/journal.pntd.0006349"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1038\/nrmicro3524","article-title":"Filovirus pathogenesis and immune evasion: Insights from Ebola virus and Marburg virus","volume":"13","author":"Messaoudi","year":"2015","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"11779","DOI":"10.1128\/JVI.01525-12","article-title":"Inclusion bodies are a site of ebolavirus replication","volume":"86","author":"Hoenen","year":"2012","journal-title":"J. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"394","DOI":"10.1038\/nature24490","article-title":"Structure and assembly of the Ebola virus nucleocapsid","volume":"551","author":"Wan","year":"2017","journal-title":"Nature"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"4275","DOI":"10.1073\/pnas.1120453109","article-title":"Structural dissection of Ebola virus and its assembly determinants using cryo-electron tomography","volume":"109","author":"Bharat","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"11165","DOI":"10.1074\/jbc.M113.461285","article-title":"Phosphorylation of Ebola virus VP30 influences the composition of the viral nucleocapsid complex: Impact on viral transcription and replication","volume":"288","author":"Biedenkopf","year":"2013","journal-title":"J. Biol. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"e02170","DOI":"10.1128\/JVI.02170-19","article-title":"The Integrity of the YxxL Motif of Ebola Virus VP24 Is Important for the Transport of Nucleocapsid\u2013Like Structures and for the Regulation of Viral RNA Synthesis","volume":"94","author":"Takamatsu","year":"2020","journal-title":"J. Virol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Dolnik, O., Kolesnikova, L., Welsch, S., Strecker, T., Schudt, G., and Becker, S. (2014). Interaction with Tsg101 is necessary for the efficient transport and release of nucleocapsids in marburg virus-infected cells. PLoS Pathog., 10.","DOI":"10.1371\/journal.ppat.1004463"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"7847","DOI":"10.1128\/JVI.00476-10","article-title":"Tsg101 is recruited by a late domain of the nucleocapsid protein to support budding of Marburg virus\u2013like particles","volume":"84","author":"Dolnik","year":"2010","journal-title":"J. Virol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e00538","DOI":"10.1128\/JVI.00538-17","article-title":"Microtubule Regulation and Function during Virus Infection","volume":"91","author":"Naghavi","year":"2017","journal-title":"J. Virol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1146\/annurev-cellbio-100818-125427","article-title":"Cell Motility and Cytokinesis: From Mysteries to Molecular Mechanisms in Five Decades","volume":"35","author":"Pollard","year":"2019","journal-title":"Annu. Rev. Cell. Dev. Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1038\/nrm4012","article-title":"Forcing cells into shape: The mechanics of actomyosin contractility","volume":"16","author":"Murrell","year":"2015","journal-title":"Nat. Rev. Mol. Cell. Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1038\/nrm2867","article-title":"A nucleator arms race: Cellular control of actin assembly","volume":"11","author":"KCampellone","year":"2010","journal-title":"Nat. Rev. Mol. Cell. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"a018226","DOI":"10.1101\/cshperspect.a018226","article-title":"Actin and Actin-Binding Proteins","volume":"18","author":"Pollard","year":"2016","journal-title":"Cold Spring Harb. Perspect Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2519","DOI":"10.1091\/mbc.e15-10-0728","article-title":"Actin-binding proteins: The long road to understanding the dynamic landscape of cellular actin networks","volume":"27","author":"Lappalainen","year":"2016","journal-title":"Mol. Biol. Cell."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1016\/j.tibs.2015.12.002","article-title":"Formins at the Junction","volume":"41","author":"Grikscheit","year":"2016","journal-title":"Trends Biochem. Sci."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"476","DOI":"10.4161\/cam.21373","article-title":"Actin on trafficking: Could actin guide directed receptor transport?","volume":"6","author":"Zech","year":"2012","journal-title":"Cell. Adh. Migr."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1042\/EBC20190015","article-title":"Actin dynamics in cell migration","volume":"63","author":"Schaks","year":"2019","journal-title":"Essays Biochem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1431","DOI":"10.1038\/ncb2353","article-title":"An actin-dependent mechanism for long-range vesicle transport","volume":"13","author":"Schuh","year":"2011","journal-title":"Nat. Cell. Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"713","DOI":"10.1038\/nrm2026","article-title":"The ARP2\/3 complex: An actin nucleator comes of age","volume":"7","author":"Goley","year":"2006","journal-title":"Nat. Rev. Mol. Cell. Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1038\/nsmb870","article-title":"Conformational changes in the Arp2\/3 complex leading to actin nucleation","volume":"12","author":"Rodal","year":"2005","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_28","first-page":"E3834","article-title":"Role and structural mechanism of WASP-triggered conformational changes in branched actin filament nucleation by Arp2\/3 complex","volume":"113","author":"Luan","year":"2016","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1038\/nrm3492","article-title":"New insights into the regulation and cellular functions of the ARP2\/3 complex","volume":"14","author":"Rotty","year":"2013","journal-title":"Nat. Rev. Mol. Cell. Biol."},{"key":"ref_30","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_31","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1126\/science.281.5373.105","article-title":"Interaction of human Arp2\/3 complex and the Listeria monocytogenes ActA protein in actin filament nucleation","volume":"281","author":"Welch","year":"1998","journal-title":"Science"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"761","DOI":"10.1111\/j.1462-5822.2004.00402.x","article-title":"A Rickettsia WASP-like protein activates the Arp2\/3 complex and mediates actin-based motility","volume":"6","author":"Jeng","year":"2004","journal-title":"Cell. Microbiol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"11390","DOI":"10.1021\/bi010486b","article-title":"Listeria protein ActA mimics WASp family proteins: It activates filament barbed end branching by Arp2\/3 complex","volume":"40","author":"Gouin","year":"2001","journal-title":"Biochemistry"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"7260","DOI":"10.1128\/JVI.00051-06","article-title":"Infection of naive target cells with virus-like particles: Implications for the function of ebola virus VP24","volume":"80","author":"Hoenen","year":"2006","journal-title":"J. Virol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"4914","DOI":"10.1128\/JVI.03257-15","article-title":"Dynamic Phosphorylation of VP30 Is Essential for Ebola Virus Life Cycle","volume":"90","author":"Biedenkopf","year":"2016","journal-title":"J. Virol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1016\/j.chembiol.2013.03.019","article-title":"Small molecules CK-666 and CK-869 inhibit actin-related protein 2\/3 complex by blocking an activating conformational change","volume":"20","author":"Hetrick","year":"2013","journal-title":"Chem. Biol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"e29795","DOI":"10.7554\/eLife.29795","article-title":"Rac1 GTPase activates the WAVE regulatory complex through two distinct binding sites","volume":"6","author":"Chen","year":"2017","journal-title":"Elife"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"790","DOI":"10.1038\/nature00859","article-title":"Mechanism of regulation of WAVE1-induced actin nucleation by Rac1 and Nck","volume":"418","author":"Eden","year":"2002","journal-title":"Nature"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2575","DOI":"10.1242\/jcs.023879","article-title":"WASP and SCAR\/WAVE proteins: The drivers of actin assembly","volume":"122","author":"Pollitt","year":"2009","journal-title":"J. Cell. Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1597","DOI":"10.1083\/jcb.109.4.1597","article-title":"Actin filaments and the growth, movement, and spread of the intracellular bacterial parasite, Listeria monocytogenes","volume":"109","author":"Tilney","year":"1989","journal-title":"J. Cell. Biol."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Mueller, J., Pfanzelter, J., Winkler, C., Narita, A., le Clainche, C., Nemethova, M., Carlier, M.F., Maeda, Y., Welch, M.D., and Ohkawa, T.C. (2014). Electron tomography and simulation of baculovirus actin comet tails support a tethered filament model of pathogen propulsion. PLoS Biol., 12.","DOI":"10.1371\/journal.pbio.1001765"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"650","DOI":"10.1016\/j.tcb.2010.08.014","article-title":"WASH, WHAMM and JMY: Regulation of Arp2\/3 complex and beyond","volume":"20","author":"Rottner","year":"2010","journal-title":"Trends Cell. Biol."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Derivery, E., Helfer, E., Henriot, V., and Gautreau, A. (2012). Actin polymerization controls the organization of WASH domains at the surface of endosomes. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0039774"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1702","DOI":"10.1080\/15548627.2015.1073434","article-title":"WHAMM links actin assembly via the Arp2\/3 complex to autophagy","volume":"11","author":"Kast","year":"2015","journal-title":"Autophagy"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1083\/jcb.201802157","article-title":"LC3 and STRAP regulate actin filament assembly by JMY during autophagosome formation","volume":"218","author":"Hu","year":"2019","journal-title":"J. Cell Biol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"8229","DOI":"10.1073\/pnas.0702454104","article-title":"A kinematic description of the trajectories of Listeria monocytogenes propelled by actin comet tails","volume":"104","author":"Shenoy","year":"2007","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Dayel, M.J., Akin, O., Landeryou, M., Risca, V., Mogilner, A., and Mullins, R.D. (2009). In silico reconstitution of actin-based symmetry breaking and motility. PLoS Biol., 7.","DOI":"10.1371\/journal.pbio.1000201"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"614","DOI":"10.1091\/mbc.e11-06-0584","article-title":"Choosing orientation: Influence of cargo geometry and ActA polarization on actin comet tails","volume":"23","author":"Lacayo","year":"2012","journal-title":"Mol. Biol. Cell."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1006\/viro.1996.0582","article-title":"Intracellular transport and processing of the Marburg virus surface protein in vertebrate and insect cells","volume":"225","author":"Becker","year":"1996","journal-title":"Virology"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"S318","DOI":"10.1093\/infdis\/jiy424","article-title":"A Fluorescently Labeled Marburg Virus Glycoprotein as a New Tool to Study Viral Transport and Assembly","volume":"218","author":"Mittler","year":"2018","journal-title":"J. Infect. Dis."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1016\/j.chom.2008.02.001","article-title":"Ebola virus matrix protein VP40 uses the COPII transport system for its intracellular transport","volume":"3","author":"Yamayoshi","year":"2008","journal-title":"Cell. Host Microbe"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"19125","DOI":"10.1038\/srep19125","article-title":"The Ebola Virus matrix protein, VP40, requires phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) for extensive oligomerization at the plasma membrane and viral egress","volume":"6","author":"Johnson","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Welsch, S., Kolesnikova, L., Kr\u00e4hling, V., Riches, J.D., Becker, S., and Briggs, J.A. (2010). Electron tomography reveals the steps in filovirus budding. PLoS Pathog, 6.","DOI":"10.1371\/journal.ppat.1000875"}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/9\/7\/1728\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:49:51Z","timestamp":1760176191000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/9\/7\/1728"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,7,19]]},"references-count":53,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2020,7]]}},"alternative-id":["cells9071728"],"URL":"https:\/\/doi.org\/10.3390\/cells9071728","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,7,19]]}}}