{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T15:59:19Z","timestamp":1791302359392,"version":"4.1.0"},"reference-count":66,"publisher":"American Chemical Society (ACS)","issue":"39","license":[{"start":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T00:00:00Z","timestamp":1790035200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100010269","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["226484\/Z\/22\/Z"],"award-info":[{"award-number":["226484\/Z\/22\/Z"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Tick-borne encephalitis virus (TBEV) invades the central nervous system (CNS) through strain-specific mechanisms that remain poorly understood. For mosquito-borne orthoflaviviruses such as dengue and yellow fever viruses, the nonstructural protein 1 (NS1) has been shown to disrupt endothelial barrier integrity by targeting tight junction proteins (TJPs), facilitating viral neuroinvasion. However, comparable mechanisms in TBEVs remain largely unexplored. Here, we investigated the potential interaction of NS1 from high (Hypr)- and low (Vs)-pathogenic TBEV strains with different blood\u2013brain barrier (BBB) TJPs, using AlphaFold3 (AF3) multimer modeling and in vitro binding assays. AF3 modeling suggested that NS1 from the highly pathogenic strain may interact with multiple TJPs, including junctional adhesion molecule A (JAM-A), a key component of the paracellular barrier. In contrast, the low-pathogenic Vs showed a more restricted interaction profile. Experimental validation using recombinant NS1 proteins revealed strain-specific binding profiles: Hypr NS1 displayed high-affinity, saturable direct binding to immobilized JAM-A (KD,app = 0.81 nM), whereas Vs NS1 showed no binding. Immunofluorescence assays on human lung epithelial (A549) and BBB (hCMEC\/D3) cells further demonstrated colocalization between Hypr NS1 and JAM-A, supporting a potential interaction. No direct binding to ZO-1, a barrier scaffold lacking an extracellular domain, was observed for either strain. This differential interaction profile may be modulated by 22 amino acids localized to the Wing and \u03b2-Ladder domains within NS1, which distinguish Hypr from Vs. Notably, despite its limited interactions with the investigated TJPs, the Vs strain is associated with slow-progressive infections that can culminate in chronic neurological disease. This highlights the need for further studies into noncanonical pathways of neuroinvasion.<\/jats:p>","DOI":"10.1021\/acsomega.6c00564","type":"journal-article","created":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T05:02:23Z","timestamp":1790053343000},"page":"58584-58598","source":"Crossref","is-referenced-by-count":0,"title":["Interactions between\nTick-Borne Encephalitis Virus\nNonstructural Protein 1 and Blood\u2013Brain Barrier Tight Junction\nProteins: Potential Clues to Strain-Specific Neuropathogenicity"],"prefix":"10.1021","volume":"11","author":[{"given":"Jarinya","family":"Chaopreecha","sequence":"first","affiliation":[{"name":"University of Leeds , , ,","place":["Leeds, U.K., LS2 9JT"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jordan","family":"Liburd","sequence":"additional","affiliation":[{"name":"University of Leeds , , ,","place":["Leeds, U.K., LS2 9JT"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shahid","family":"Khalid","sequence":"additional","affiliation":[{"name":"University of Leeds , , ,","place":["Leeds, U.K., LS2 9JT"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Songeziwe","family":"Ntsimango","sequence":"additional","affiliation":[{"name":"University of the Witwatersrand , , PO Wits , ,","place":["Johannesburg, South Africa, 2050"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ian M.","family":"Jones","sequence":"additional","affiliation":[{"name":"University of Reading , , ,","place":["Reading, U.K., RG6 6UR"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer H.","family":"Tomlinson","sequence":"additional","affiliation":[{"name":"University of Leeds , , ,","place":["Leeds, U.K., LS2 9JT"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2906-5513","authenticated-orcid":true,"given":"Niluka","family":"Goonawardane","sequence":"additional","affiliation":[{"name":"University of Leeds , , ,","place":["Leeds, U.K., LS2 9JT"]},{"name":"University of Oxford , , Life and Mind Building, South Parks Road , ,","place":["Oxford, U.K., OX1 3EL"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,22]]},"reference":[{"key":"2026100610495578500_cit1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/S0065-3527(03)59001-7","article-title":"Taxonomy of the virus family Flaviviridae","volume":"59","author":"Calisher","year":"2003","journal-title":"Adv. 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Neurol."},{"issue":"7","key":"2026100610495578500_cit5","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0004828","article-title":"Dengue Virus Nonstructural Protein\n1 Induces Vascular Leakage through Macrophage Migration Inhibitory\nFactor and Autophagy","volume":"10","author":"Chen","year":"2016","journal-title":"PLoS Negl. Trop. Dis."},{"issue":"6","key":"2026100610495578500_cit6","doi-asserted-by":"publisher","first-page":"615","DOI":"10.3390\/pathogens11060615","article-title":"Flavivirus NS1 Triggers\nTissue-Specific Disassembly of Intercellular Junctions Leading to\nBarrier Dysfunction and Vascular Leak in a GSK-3beta-Dependent Manner","volume":"11","author":"Puerta-Guardo","year":"2022","journal-title":"Pathogens"},{"issue":"5","key":"2026100610495578500_cit7","doi-asserted-by":"publisher","first-page":"2342","DOI":"10.3390\/ijms26052342","article-title":"Impact of Tick-Borne\nOrthoflaviviruses Infection on Compact Human Brain Endothelial Barrier","volume":"26","author":"Schweitzer","year":"2025","journal-title":"Int. J. Mol. 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Virol."},{"issue":"6","key":"2026100610495578500_cit26","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0158105","article-title":"Tick-Borne\nEncephalitis Virus Structural Proteins\nAre the Primary Viral Determinants of Non-Viraemic Transmission between\nTicks whereas Non-Structural Proteins Affect Cytotoxicity","volume":"11","author":"Khasnatinov","year":"2016","journal-title":"PLoS One"},{"issue":"12","key":"2026100610495578500_cit27","doi-asserted-by":"publisher","first-page":"9608","DOI":"10.1128\/jvi.71.12.9608-9617.1997","article-title":"trans-Complementation\nof yellow fever virus NS1 reveals\na role in early RNA replication","volume":"71","author":"Lindenbach","year":"1997","journal-title":"J. Virol."},{"issue":"7","key":"2026100610495578500_cit28","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1005738","article-title":"Dengue Virus NS1 Disrupts the Endothelial\nGlycocalyx, Leading to Hyperpermeability","volume":"12","author":"Puerta-Guardo","year":"2016","journal-title":"PLoS\nPathog."},{"issue":"8016","key":"2026100610495578500_cit29","doi-asserted-by":"publisher","first-page":"493","DOI":"10.1038\/s41586-024-07487-w","article-title":"Accurate\nstructure prediction of biomolecular\ninteractions with AlphaFold 3","volume":"630","author":"Abramson","year":"2024","journal-title":"Nature"},{"key":"2026100610495578500_cit30","doi-asserted-by":"publisher","first-page":"1566486","DOI":"10.3389\/fbinf.2025.1566486","article-title":"Using AlphaFold-Multimer to study novel protein-protein interactions\nof predation essential hypothetical proteins in Bdellovibrio","volume":"5","author":"Abulude","year":"2025","journal-title":"Front. Bioinform."},{"issue":"1","key":"2026100610495578500_cit31","doi-asserted-by":"publisher","first-page":"78","DOI":"10.1186\/s12896-018-0486-2","article-title":"Association\nof high pressure and alkaline condition for solubilization of inclusion\nbodies and refolding of the NS1 protein from zika virus","volume":"18","author":"Rosa da Silva","year":"2018","journal-title":"BMC Biotechnol."},{"key":"2026100610495578500_cit32","doi-asserted-by":"publisher","first-page":"72","DOI":"10.1016\/j.virol.2022.06.005","article-title":"Structural dynamics\nof Zika virus\nNS1 via a reductionist approach reveal the disordered nature of its\nbeta-roll domain in isolation","volume":"573","author":"Kapuganti","year":"2022","journal-title":"Virology"},{"key":"2026100610495578500_cit33","unstructured":"The PyMOL Molecular\nGraphics System; DeLano Scientific: San Carlos, CA,\nUSA, 2002. http:\/\/www.pymol.org."},{"issue":"W1","key":"2026100610495578500_cit34","doi-asserted-by":"publisher","first-page":"W365","DOI":"10.1093\/nar\/gkx407","article-title":"HDOCK:\na web server for protein-protein and protein-DNA\/RNA docking based\non a hybrid strategy","volume":"45","author":"Yan","year":"2017","journal-title":"Nucleic Acids Res."},{"key":"2026100610495578500_cit35","doi-asserted-by":"publisher","first-page":"94","DOI":"10.1016\/j.bbrc.2021.03.100","article-title":"SARS-CoV-2 spike protein\ninteractions with amyloidogenic proteins:\nPotential clues to neurodegeneration","volume":"554","author":"Idrees","year":"2021","journal-title":"Biochem.\nBiophys. Res. Commun."},{"issue":"18","key":"2026100610495578500_cit36","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.00818-22","article-title":"Identification\nof\nHost Factors Differentially Induced by Clinically Diverse Strains\nof Tick-Borne Encephalitis Virus","volume":"96","author":"Goonawardane","year":"2022","journal-title":"J. Virol."},{"issue":"2","key":"2026100610495578500_cit37","doi-asserted-by":"publisher","first-page":"1497","DOI":"10.1128\/jvi.71.2.1497-1505.1997","article-title":"Subgenomic replicons of the flavivirus Kunjin: construction\nand applications","volume":"71","author":"Khromykh","year":"1997","journal-title":"J. Virol."},{"issue":"7","key":"2026100610495578500_cit38","doi-asserted-by":"publisher","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: an open-source platform for biological-image\nanalysis","volume":"9","author":"Schindelin","year":"2012","journal-title":"Nat. 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Commun."},{"issue":"10","key":"2026100610495578500_cit42","doi-asserted-by":"publisher","first-page":"951","DOI":"10.1056\/NEJMoa1600651","article-title":"Zika Virus Associated\nwith Microcephaly","volume":"374","author":"Mlakar","year":"2016","journal-title":"N. Engl. J. Med."},{"issue":"6365","key":"2026100610495578500_cit43","doi-asserted-by":"publisher","first-page":"933","DOI":"10.1126\/science.aam7120","article-title":"A single mutation in the prM protein of Zika virus\ncontributes to fetal microcephaly","volume":"358","author":"Yuan","year":"2017","journal-title":"Science"},{"issue":"1","key":"2026100610495578500_cit44","doi-asserted-by":"publisher","first-page":"916","DOI":"10.1038\/s41467-021-21199-z","article-title":"Recent African strains of Zika virus display higher\ntransmissibility and fetal pathogenicity than Asian strains","volume":"12","author":"Aubry","year":"2021","journal-title":"Nat. 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Virol."},{"issue":"2","key":"2026100610495578500_cit46","doi-asserted-by":"publisher","first-page":"49","DOI":"10.3390\/v8020049","article-title":"In Vitro and\nin Vivo Evaluation of Mutations in the NS Region of Lineage 2 West\nNile Virus Associated with Neuroinvasiveness in a Mammalian Model","volume":"8","author":"Szentpali-Gavaller","year":"2016","journal-title":"Viruses"},{"issue":"5","key":"2026100610495578500_cit47","doi-asserted-by":"publisher","first-page":"929","DOI":"10.3201\/eid3105.240893","article-title":"Powassan and Eastern\nEquine Encephalitis Virus Seroprevalence in Endemic Areas, United\nStates, 2019-2020","volume":"31","author":"Padda","year":"2025","journal-title":"Emerg. Infect. Dis."},{"issue":"12","key":"2026100610495578500_cit48","doi-asserted-by":"publisher","first-page":"6290","DOI":"10.1007\/s12035-021-02548-x","article-title":"Zika Virus NS1 Suppresses VE-Cadherin and Claudin-5\nvia hsa-miR-101-3p in Human Brain Microvascular Endothelial Cells","volume":"58","author":"Bhardwaj","year":"2021","journal-title":"Mol. Neurobiol."},{"issue":"1","key":"2026100610495578500_cit49","first-page":"1","article-title":"Significance of ZO-1, an Intercellular Adhesion\nMolecule, as a Prognostic Marker in Lung Adenocarcinoma","volume":"49","author":"Tsuboi","year":"2024","journal-title":"Tokai J. Exp Clin Med."},{"key":"2026100610495578500_cit50","doi-asserted-by":"publisher","first-page":"110659","DOI":"10.1016\/j.cellsig.2023.110659","article-title":"Zika virus NS1 suppresses VE-cadherin via hsa-miR-29b-3p\/DNMT3b\/MMP-9\npathway in human brain microvascular endothelial cells","volume":"106","author":"Bhardwaj","year":"2023","journal-title":"Cell. 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Virol."},{"issue":"6525","key":"2026100610495578500_cit57","doi-asserted-by":"publisher","first-page":"190","DOI":"10.1126\/science.abb9425","article-title":"A broadly protective\nantibody that targets\nthe flavivirus NS1 protein","volume":"371","author":"Modhiran","year":"2021","journal-title":"Science"},{"issue":"11","key":"2026100610495578500_cit58","doi-asserted-by":"publisher","first-page":"793","DOI":"10.1590\/0074-02760170299","article-title":"Tracing the origin of the NS1 A188V substitution responsible\nfor recent enhancement of Zika virus Asian genotype infectivity","volume":"112","author":"Delatorre","year":"2017","journal-title":"Mem. Inst. Oswaldo Cruz."},{"issue":"12","key":"2026100610495578500_cit59","doi-asserted-by":"publisher","first-page":"6125","DOI":"10.1128\/JVI.02617-08","article-title":"West nile virus\ncapsid degradation of claudin proteins disrupts epithelial barrier\nfunction","volume":"83","author":"Medigeshi","year":"2009","journal-title":"J. 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