{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T23:27:28Z","timestamp":1790724448715,"version":"4.1.0"},"reference-count":28,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2017,11,2]],"date-time":"2017-11-02T00:00:00Z","timestamp":1509580800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Health and Medical Research Council of Australia","award":["APP1045188"],"award-info":[{"award-number":["APP1045188"]}]},{"name":"National Health and Medical Research Council of Australia","award":["APP1059794"],"award-info":[{"award-number":["APP1059794"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>West Nile virus (WNV) is a neurotropic flavivirus that can cause encephalitis in mammalian and avian hosts. In America, the virulent WNV strain (NY99) is causing yearly outbreaks of encephalitis in humans and horses, while in Australia the less virulent Kunjin strain of WNV strain has not been associated with significant disease outbreaks until a recent 2011 large outbreak in horses (but not in humans) caused by NSW2011 strain. Using chimeric viruses between NY99 and NSW2011 strains we previously identified a role for the non-structural proteins of NY99 strain and especially the NS3 protein, in enhanced virus replication in type I interferon response-competent cells and increased virulence in mice. To further define the role of NY99 NS3 protein in inhibition of type I interferon response, we have generated and characterised additional chimeric viruses containing the protease or the helicase domains of NY99 NS3 on the background of the NSW2011 strain. The results identified the role for the helicase but not the protease domain of NS3 protein in the inhibition of type I interferon signalling and showed that helicase domain of the more virulent NY99 strain performs this function more efficiently than helicase domain of the less virulent NSW2011 strain. Further analysis with individual amino acid mutants identified two amino acid residues in the helicase domain primarily responsible for this difference. Using chimeric replicons, we also showed that the inhibition of type I interferon (IFN) signalling was independent of other known functions of NS3 in RNA replication and assembly of virus particles.<\/jats:p>","DOI":"10.3390\/v9110326","type":"journal-article","created":{"date-parts":[[2017,11,3]],"date-time":"2017-11-03T04:43:13Z","timestamp":1509684193000},"page":"326","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["Helicase Domain of West Nile Virus NS3 Protein Plays a Role in Inhibition of Type I Interferon Signalling"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1683-3555","authenticated-orcid":false,"given":"Yin","family":"Setoh","sequence":"first","affiliation":[{"name":"Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, St. Lucia, QLD 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Parthiban","family":"Periasamy","sequence":"additional","affiliation":[{"name":"Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, St. Lucia, QLD 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7488-5441","authenticated-orcid":false,"given":"Nias","family":"Peng","sequence":"additional","affiliation":[{"name":"Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, St. Lucia, QLD 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alberto","family":"Amarilla","sequence":"additional","affiliation":[{"name":"Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, St. Lucia, QLD 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3258-0508","authenticated-orcid":false,"given":"Andrii","family":"Slonchak","sequence":"additional","affiliation":[{"name":"Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, St. Lucia, QLD 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander","family":"Khromykh","sequence":"additional","affiliation":[{"name":"Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, St. Lucia, QLD 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2017,11,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"792","DOI":"10.3201\/eid1805.111720","article-title":"Characterization of virulent west nile virus kunjin strain, australia, 2011","volume":"18","author":"Frost","year":"2012","journal-title":"Emerg. Infect. Dis."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1297","DOI":"10.1099\/vir.0.000069","article-title":"Systematic analysis of viral genes responsible for differential virulence between american and australian west nile virus strains","volume":"96","author":"Setoh","year":"2015","journal-title":"J. Gen. Virol."},{"key":"ref_3","unstructured":"Knipe, D.M., and Howley, P.M. (2007). Flaviviridae: The viruses and their replication. Fields Virology, Lippincott-Raven Publishers. [5th ed.]."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1551","DOI":"10.1099\/vir.0.000097","article-title":"Post-translational regulation and modifications of flavivirus structural proteins","volume":"96","author":"Roby","year":"2015","journal-title":"J. Gen. Virol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1146\/annurev.biochem.76.052305.115300","article-title":"Structure and mechanism of helicases and nucleic acid translocases","volume":"76","author":"Singleton","year":"2007","journal-title":"Annu. Rev. Biochem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"444","DOI":"10.1016\/j.jmb.2007.06.055","article-title":"Crystal structure and activity of kunjin virus NS3 helicase; protease and helicase domain assembly in the full length NS3 protein","volume":"372","author":"Mastrangelo","year":"2007","journal-title":"J. Mol. Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"3342","DOI":"10.1128\/JVI.02447-07","article-title":"Yellow fever virus NS3 plays an essential role in virus assembly independent of its known enzymatic functions","volume":"82","author":"Patkar","year":"2008","journal-title":"J. Virol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"11255","DOI":"10.1128\/JVI.01559-06","article-title":"Translation of the flavivirus kunjin NS3 gene in cis but not its rna sequence or secondary structure is essential for efficient rna packaging","volume":"80","author":"Pijlman","year":"2006","journal-title":"J. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Aguirre, S., Maestre, A.M., Pagni, S., Patel, J.R., Savage, T., Gutman, D., Maringer, K., Bernal-Rubio, D., Shabman, R.S., and Simon, V. (2012). Denv inhibits type i ifn production in infected cells by cleaving human sting. PLoS Pathog., 8.","DOI":"10.1371\/journal.ppat.1002934"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"De Borba, L., Strottmann, D.M., de Noronha, L., Mason, P.W., and Dos Santos, C.N. (2012). Synergistic interactions between the NS3(hel) and e proteins contribute to the virulence of dengue virus type 1. PLoS Negl. Trop. Dis., 6.","DOI":"10.1371\/journal.pntd.0001624"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1111\/cei.12701","article-title":"Single point mutations in the helicase domain of the NS3 protein enhance dengue virus replicative capacity in human monocyte-derived dendritic cells and circumvent the type i interferon response","volume":"183","author":"Silveira","year":"2016","journal-title":"Clin. Exp. Immunol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Langevin, S.A., Bowen, R.A., Reisen, W.K., Andrade, C.C., Ramey, W.N., Maharaj, P.D., Anishchenko, M., Kenney, J.L., Duggal, N.K., and Romo, H. (2014). Host competence and helicase activity differences exhibited by west nile viral variants expressing NS3-249 amino acid polymorphisms. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0100802"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1162","DOI":"10.1038\/ng2097","article-title":"A single positively selected west nile viral mutation confers increased virogenesis in american crows","volume":"39","author":"Brault","year":"2007","journal-title":"Nat. Genet."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1016\/j.virol.2011.03.008","article-title":"Virulence determinants between new york 99 and kunjin strains of west nile virus","volume":"414","author":"Audsley","year":"2011","journal-title":"Virology"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2367","DOI":"10.1128\/JVI.03162-12","article-title":"A novel bacterium-free method for generation of flavivirus infectious DNA by circular polymerase extension reaction allows accurate recapitulation of viral heterogeneity","volume":"87","author":"Edmonds","year":"2013","journal-title":"J. Virol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1497","DOI":"10.1128\/jvi.71.2.1497-1505.1997","article-title":"Subgenomic replicons of the flavivirus kunjin: Construction and applications","volume":"71","author":"Khromykh","year":"1997","journal-title":"J. Virol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1128\/JVI.78.1.531-538.2004","article-title":"Tetracycline-inducible packaging cell line for production of flavivirus replicon particles","volume":"78","author":"Harvey","year":"2004","journal-title":"J. Virol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1175","DOI":"10.1099\/vir.0.82609-0","article-title":"In situ reactions of monoclonal antibodies with a viable mutant of murray valley encephalitis virus reveal an absence of dimeric ns1 protein","volume":"88","author":"Clark","year":"2007","journal-title":"J. Gen. Virol."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Setoh, Y.X., Prow, N.A., Peng, N., Hugo, L.E., Devine, G., Hazlewood, J.E., Suhrbier, A., and Khromykh, A.A. (2017). De novo generation and characterization of new zika virus isolate using sequence data from a microcephaly case. mSphere, 2.","DOI":"10.1128\/mSphereDirect.00190-17"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"44642","DOI":"10.1038\/srep44642","article-title":"Chimeric viruses between rocio and west nile: The role for rocio prm-e proteins in virulence and inhibition of interferon-alpha\/beta signaling","volume":"7","author":"Amarilla","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"3503","DOI":"10.1128\/JVI.01161-09","article-title":"The NS5 protein of the virulent west nile virus NY99 strain is a potent antagonist of type i interferon-mediated JAK-STAT signaling","volume":"84","author":"Boer","year":"2010","journal-title":"J. Virol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Lazear, H.M., Lancaster, A., Wilkins, C., Suthar, M.S., Huang, A., Vick, S.C., Clepper, L., Thackray, L., Brassil, M.M., and Virgin, H.W. (2013). IRF-3, IRF-5 and IRF-7 coordinately regulate the type I IFN response in myeloid dendritic cells downstream of MAVS signaling. PLoS Pathog., 9.","DOI":"10.1371\/annotation\/4de7ddfd-52df-4f87-8ca4-d48afe646ca8"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Suthar, M.S., Ma, D.Y., Thomas, S., Lund, J.M., Zhang, N., Daffis, S., Rudensky, A.Y., Bevan, M.J., Clark, E.A., and Kaja, M.K. (2010). IPS-1 is essential for the control of west nile virus infection and immunity. PLoS Pathog., 6.","DOI":"10.1371\/journal.ppat.1000757"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/S0959-440X(02)00298-1","article-title":"Helicase structure and mechanism","volume":"12","author":"Caruthers","year":"2002","journal-title":"Curr. Opin. Struct. Biol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"10268","DOI":"10.1128\/JVI.79.16.10268-10277.2005","article-title":"Structure of the flavivirus helicase: Implications for catalytic activity, protein interactions and proteolytic processing","volume":"79","author":"Wu","year":"2005","journal-title":"J. Virol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1146\/annurev.ento.53.103106.093258","article-title":"A global perspective on the epidemiology of west nile virus","volume":"53","author":"Kramer","year":"2008","journal-title":"Annu. Rev. Entomol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"6255","DOI":"10.3390\/ijerph10126255","article-title":"The changing epidemiology of kunjin virus in australia","volume":"10","author":"Prow","year":"2013","journal-title":"Int. J. Environ. Res. Public Health"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1111\/avj.12018","article-title":"Descriptive overview of the 2011 epidemic of arboviral disease in horses in australia","volume":"91","author":"Roche","year":"2013","journal-title":"Aust. Vet. J."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/9\/11\/326\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T18:47:52Z","timestamp":1760208472000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/9\/11\/326"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,11,2]]},"references-count":28,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2017,11]]}},"alternative-id":["v9110326"],"URL":"https:\/\/doi.org\/10.3390\/v9110326","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,11,2]]}}}