{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,11]],"date-time":"2026-02-11T14:14:15Z","timestamp":1770819255561,"version":"3.50.1"},"reference-count":40,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2020,3,13]],"date-time":"2020-03-13T00:00:00Z","timestamp":1584057600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100010203","name":"Agriculture Research System of China","doi-asserted-by":"publisher","award":["grant no. CARS-37"],"award-info":[{"award-number":["grant no. CARS-37"]}],"id":[{"id":"10.13039\/501100010203","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2017YFD0500904"],"award-info":[{"award-number":["2017YFD0500904"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2018YFD0501600"],"award-info":[{"award-number":["2018YFD0501600"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Bovine herpesvirus1 (BoHV-1) is a major bovine pathogen. Despite several vaccines being available to prevent viral infection, outbreaks are frequent and cause important economic consequences worldwide. The development of new antiviral drugs is therefore highly desirable. In this context, viral genome replication represents a potential target for therapeutic intervention. BoHV-1 genome is a dsDNA molecule whose replication takes place in the nuclei of infected cells and is mediated by a viral encoded DNA polymerase holoenzyme. Here, we studied the physical interaction and subcellular localization of BoHV-1 DNA polymerase subunits in cells for the first time. By means of co-immunoprecipitation and confocal laser scanning microscopy (CLSM) experiments, we could show that the processivity factor of the DNA polymerase pUL42 is capable of being autonomously transported into the nucleus, whereas the catalytic subunit pUL30 is not. Accordingly, a putative classic NLS (cNLS) was identified on pUL42 but not on pUL30. Importantly, both proteins could interact in the absence of other viral proteins and their co-expression resulted in accumulation of UL30 to the cell nucleus. Treatment of cells with Ivermectin, an anti-parasitic drug which has been recently identified as an inhibitor of importin \u03b1\/\u03b2-dependent nuclear transport, reduced UL42 nuclear import and specifically reduced BoHV-1 replication in a dose-dependent manner, while virus attachment and entry into cells were not affected. Therefore, this study provides a new option of antiviral therapy for BoHV-1 infection with Ivermectin.<\/jats:p>","DOI":"10.3390\/microorganisms8030409","type":"journal-article","created":{"date-parts":[[2020,3,17]],"date-time":"2020-03-17T09:27:41Z","timestamp":1584437261000},"page":"409","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":45,"title":["Ivermectin Inhibits Bovine Herpesvirus 1 DNA Polymerase Nuclear Import and Interferes With Viral Replication"],"prefix":"10.3390","volume":"8","author":[{"given":"Sohail","family":"Raza","sequence":"first","affiliation":[{"name":"The State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"Department of Microbiology, University of Veterinary and Animal Sciences, Lahore 54000, Pakistan"},{"name":"College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Farzana","family":"Shahin","sequence":"additional","affiliation":[{"name":"The State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wenjun","family":"Zhai","sequence":"additional","affiliation":[{"name":"The State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hanxiong","family":"Li","sequence":"additional","affiliation":[{"name":"The State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8177-3616","authenticated-orcid":false,"given":"Gualtiero","family":"Alvisi","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, University of Padua, 35121 Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kui","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA 70803, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xi","family":"Chen","sequence":"additional","affiliation":[{"name":"The State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yingyu","family":"Chen","sequence":"additional","affiliation":[{"name":"The State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"Key Laboratory of Development of Veterinary Diagnostic Products, Key Laboratory of Ruminant Bio-Products of Ministry of Agriculture and Rural Affairs, Wuhan 430070, China"},{"name":"Hubei International Scientific and Technological Cooperation Base of Veterinary Epidemiology, International Research Center for Animal Disease, Ministry of Science and Technology of the People\u2019s Republic of China, Wuhan 430070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jianguo","family":"Chen","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Changmin","family":"Hu","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Huanchun","family":"Chen","sequence":"additional","affiliation":[{"name":"The State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"Key Laboratory of Development of Veterinary Diagnostic Products, Key Laboratory of Ruminant Bio-Products of Ministry of Agriculture and Rural Affairs, Wuhan 430070, China"},{"name":"Hubei International Scientific and Technological Cooperation Base of Veterinary Epidemiology, International Research Center for Animal Disease, Ministry of Science and Technology of the People\u2019s Republic of China, Wuhan 430070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aizhen","family":"Guo","sequence":"additional","affiliation":[{"name":"The State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"Key Laboratory of Development of Veterinary Diagnostic Products, Key Laboratory of Ruminant Bio-Products of Ministry of Agriculture and Rural Affairs, Wuhan 430070, China"},{"name":"Hubei International Scientific and Technological Cooperation Base of Veterinary Epidemiology, International Research Center for Animal Disease, Ministry of Science and Technology of the People\u2019s Republic of China, Wuhan 430070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,3,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"12235","DOI":"10.18632\/oncotarget.7771","article-title":"A bovine herpesvirus 1 pUL51 deletion mutant shows impaired viral growth in vitro and reduced virulence in rabbits","volume":"7","author":"Raza","year":"2016","journal-title":"Oncotarget"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/j.rvsc.2016.10.004","article-title":"Effectiveness of a BHV-1\/BEFV bivalent vaccine against bovine herpesvirus type 1 infection in cattle","volume":"109","author":"Chung","year":"2016","journal-title":"Res. Vet. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Workman, A., Zhu, L., Keel, B.N., Smith, T.P.L., and Jones, C. (2018). The Wnt Signaling Pathway Is Differentially Expressed during the Bovine Herpesvirus 1 Latency-Reactivation Cycle: Evidence That Two Protein Kinases Associated with Neuronal Survival, Akt3 and BMPR2, Are Expressed at Higher Levels during Latency. J. Virol., 92.","DOI":"10.1128\/JVI.01937-17"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"El-Mayet, F.S., Sawant, L., Thunuguntla, P., and Jones, C. (2017). Combinatorial Effects of the Glucocorticoid Receptor and Kruppel-Like Transcription Factor 15 on Bovine Herpesvirus 1 Transcription and Productive Infection. J. Virol., 91.","DOI":"10.1128\/JVI.00904-17"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Sawant, L., Kook, I., Vogel, J.L., Kristie, T.M., and Jones, C. (2018). The Cellular Coactivator HCF-1 Is Required for Glucocorticoid Receptor-Mediated Transcription of Bovine Herpesvirus 1 Immediate Early Genes. J. Virol., 92.","DOI":"10.1128\/JVI.00987-18"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Zhu, L., Workman, A., and Jones, C. (2017). Potential Role for a beta-Catenin Coactivator (High-Mobility Group AT-Hook 1 Protein) during the Latency-Reactivation Cycle of Bovine Herpesvirus 1. J. Virol., 91.","DOI":"10.1128\/JVI.02132-16"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1051\/vetres:2006059","article-title":"Bovine herpesvirus 1 infection and infectious bovine rhinotracheitis","volume":"38","author":"Muylkens","year":"2007","journal-title":"Vet. Res."},{"key":"ref_8","first-page":"132","article-title":"Novel inhibitors of human CMV","volume":"9","author":"Andrei","year":"2008","journal-title":"Curr. Opin. Investig. Drugs"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1249","DOI":"10.1002\/med.21281","article-title":"A cutting-edge view on the current state of antiviral drug development","volume":"33","year":"2013","journal-title":"Med. Res. Rev."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2210","DOI":"10.3390\/v5092210","article-title":"Regulated transport into the nucleus of herpesviridae DNA replication core proteins","volume":"5","author":"Alvisi","year":"2013","journal-title":"Viruses"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"6979","DOI":"10.1128\/jvi.67.12.6979-6988.1993","article-title":"Eleven loci encoding trans-acting factors are required for transient complementation of human cytomegalovirus oriLyt-dependent DNA replication","volume":"67","author":"Pari","year":"1993","journal-title":"J. Virol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1016\/S0960-9822(95)00079-0","article-title":"Two different subunits of importin cooperate to recognize nuclear localization signals and bind them to the nuclear envelope","volume":"5","author":"Gorlich","year":"1995","journal-title":"Curr. Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1016\/j.cell.2007.10.015","article-title":"SnapShot: Nuclear transport","volume":"131","author":"Tran","year":"2007","journal-title":"Cell"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1038\/311033a0","article-title":"Sequence requirements for nuclear location of simian virus 40 large-T antigen","volume":"311","author":"Kalderon","year":"1984","journal-title":"Nature"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"452","DOI":"10.1038\/nsmb1229","article-title":"Structure-based design of a pathway-specific nuclear import inhibitor","volume":"14","author":"Cansizoglu","year":"2007","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1002","DOI":"10.1111\/j.1600-0854.2005.00331.x","article-title":"A protein kinase CK2 site flanking the nuclear targeting signal enhances nuclear transport of human cytomegalovirus ppUL44","volume":"6","author":"Alvisi","year":"2005","journal-title":"Traffic"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1322","DOI":"10.1111\/j.1600-0854.2006.00477.x","article-title":"Human cytomegalovirus DNA polymerase catalytic subunit pUL54 possesses independently acting nuclear localization and ppUL44 binding motifs","volume":"7","author":"Alvisi","year":"2006","journal-title":"Traffic"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"9155","DOI":"10.1021\/bi7002394","article-title":"An importin alpha\/beta-recognized bipartite nuclear localization signal mediates targeting of the human herpes simplex virus type 1 DNA polymerase catalytic subunit pUL30 to the nucleus","volume":"46","author":"Alvisi","year":"2007","journal-title":"Biochemistry"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"13764","DOI":"10.1021\/bi800869y","article-title":"Nuclear import of HSV-1 DNA polymerase processivity factor UL42 is mediated by a C-terminally located bipartite nuclear localization signal","volume":"47","author":"Alvisi","year":"2008","journal-title":"Biochemistry"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1016\/j.virol.2005.06.017","article-title":"Processivity factor of KSHV contains a nuclear localization signal and binding domains for transporting viral DNA polymerase into the nucleus","volume":"340","author":"Chen","year":"2005","journal-title":"Virology"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"6482","DOI":"10.1128\/JVI.03428-12","article-title":"Nuclear Transport of Epstein-Barr Virus DNA Polymerase is dependent on the BMRF1 Polymerase Processivity Factor and Molecular Chaperone Hsp90","volume":"87","author":"Kawashima","year":"2013","journal-title":"J. Virol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/j.antiviral.2018.09.010","article-title":"Ivermectin inhibits DNA polymerase UL42 of pseudorabies virus entrance into the nucleus and proliferation of the virus in vitro and vivo","volume":"159","author":"Lv","year":"2018","journal-title":"Antivir. Res."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Yang, S.N.Y., Atkinson, S.C., Wang, C., Lee, A., Bogoyevitch, M.A., Borg, N.A., and Jans, D.A. (2020). The broad spectrum antiviral ivermectin targets the host nuclear transport importin alpha\/beta1 heterodimer. Antivir. Res., 104760.","DOI":"10.1016\/j.antiviral.2020.104760"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"9567","DOI":"10.1128\/JVI.00669-09","article-title":"The flexible loop of the human cytomegalovirus DNA polymerase processivity factor ppUL44 is required for efficient DNA binding and replication in cells","volume":"83","author":"Alvisi","year":"2009","journal-title":"J. Virol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1016\/j.virol.2011.06.015","article-title":"Multiple phosphorylation sites at the C-terminus regulate nuclear import of HCMV DNA polymerase processivity factor ppUL44","volume":"417","author":"Alvisi","year":"2011","journal-title":"Virology"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"478","DOI":"10.1074\/jbc.M807017200","article-title":"Six classes of nuclear localization signals specific to different binding grooves of importin alpha","volume":"284","author":"Kosugi","year":"2009","journal-title":"J. Biol. Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1455","DOI":"10.1042\/BCJ20170897","article-title":"Intersectin goes nuclear: Secret life of an endocytic protein","volume":"475","author":"Alvisi","year":"2018","journal-title":"Biochem. J."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"94462","DOI":"10.18632\/oncotarget.21776","article-title":"Bovine herpesvirus 1 tegument protein UL21 plays critical roles in viral secondary envelopment and cell-to-cell spreading","volume":"8","author":"Shahin","year":"2017","journal-title":"Oncotarget"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1016\/j.antiviral.2015.03.009","article-title":"Antiviral activity of theaflavin digallate against herpes simplex virus type 1","volume":"118","author":"Prince","year":"2015","journal-title":"Antivir. Res."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Cantatore, A., Randall, S.D., Traum, D., and Adams, S.D. (2013). Effect of black tea extract on herpes simplex virus-1 infection of cultured cells. BMC Complement.Altern. Med., 13.","DOI":"10.1186\/1472-6882-13-139"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1017\/S1466252313000042","article-title":"Immunity to bovine herpesvirus 1: I. Viral lifecycle and innate immunity","volume":"14","author":"Levings","year":"2013","journal-title":"Anim. Health Res. Rev."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Xu, T.L., Han, Y., Liu, W., Pang, X.Y., Zheng, B., Zhang, Y., and Zhou, X.N. (2018). Antivirus effectiveness of ivermectin on dengue virus type 2 in Aedes albopictus. PLoS Negl. Trop. Dis., 12.","DOI":"10.1371\/journal.pntd.0006934"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"11214","DOI":"10.1128\/JVI.01737-13","article-title":"Bovine herpesvirus 1 regulatory proteins bICP0 and VP16 are readily detected in trigeminal ganglionic neurons expressing the glucocorticoid receptor during the early stages of reactivation from latency","volume":"87","author":"Kook","year":"2013","journal-title":"J. Virol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"5976","DOI":"10.1128\/jvi.64.12.5976-5987.1990","article-title":"The Herpes-Simplex Virus Type-1 Ul42 Gene-Product\u2014A Subunit of DNA-Polymerase That Functions to Increase Processivity","volume":"64","author":"Gottlieb","year":"1990","journal-title":"J. Virol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"15773","DOI":"10.1073\/pnas.1520952112","article-title":"Profile of William C. Campbell, Satoshi Omura, and Youyou Tu, 2015 Nobel Laureates in Physiology or Medicine","volume":"112","author":"Wells","year":"2015","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1016\/j.rvsc.2010.09.016","article-title":"Ivermectin effects on motor coordination and contractions of isolated rat diaphragm","volume":"91","author":"Trailovic","year":"2011","journal-title":"Res. Vet. Sci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"851","DOI":"10.1042\/BJ20120150","article-title":"Ivermectin is a specific inhibitor of importin alpha\/beta-mediated nuclear import able to inhibit replication of HIV-1 and dengue virus","volume":"443","author":"Wagstaff","year":"2012","journal-title":"Biochem. J."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1016\/j.chom.2016.07.004","article-title":"A Screen of FDA-Approved Drugs for Inhibitors of Zika Virus Infection","volume":"20","author":"Barrows","year":"2016","journal-title":"Cell Host Microb."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1884","DOI":"10.1093\/jac\/dks147","article-title":"Ivermectin is a potent inhibitor of flavivirus replication specifically targeting NS3 helicase activity: New prospects for an old drug","volume":"67","author":"Mastrangelo","year":"2012","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"328","DOI":"10.4149\/av_2016_03_328","article-title":"The synergistic antiviral effects of GSH in combination with acyclovir against BoHV-1 infection in vitro","volume":"60","author":"Yuan","year":"2016","journal-title":"Acta Virol."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/8\/3\/409\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:06:46Z","timestamp":1760173606000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/8\/3\/409"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,3,13]]},"references-count":40,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2020,3]]}},"alternative-id":["microorganisms8030409"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms8030409","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,3,13]]}}}