{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T19:29:04Z","timestamp":1791919744371,"version":"4.3.4"},"reference-count":57,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2024,8,16]],"date-time":"2024-08-16T00:00:00Z","timestamp":1723766400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Centers of Biomedical Research Excellence","award":["P30GM114737"],"award-info":[{"award-number":["P30GM114737"]}]},{"name":"Centers of Biomedical Research Excellence","award":["U54GM138062"],"award-info":[{"award-number":["U54GM138062"]}]},{"name":"Center for Pacific Innovations, Knowledge, and Opportunities","award":["P30GM114737"],"award-info":[{"award-number":["P30GM114737"]}]},{"name":"Center for Pacific Innovations, Knowledge, and Opportunities","award":["U54GM138062"],"award-info":[{"award-number":["U54GM138062"]}]},{"name":"National Institute for General Medical Sciences (NIGMS)","award":["P30GM114737"],"award-info":[{"award-number":["P30GM114737"]}]},{"name":"National Institute for General Medical Sciences (NIGMS)","award":["U54GM138062"],"award-info":[{"award-number":["U54GM138062"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>West Nile virus (WNV) nonstructural protein 5 (NS5) possesses multiple enzymatic domains essential for viral RNA replication. During infection, NS5 predominantly localizes to unique replication organelles (ROs) at the rough endoplasmic reticulum (RER), known as vesicle packets (VPs) and convoluted membranes (CMs), with a portion of NS5 accumulating in the nucleus. NS5 is a soluble protein that must be in the VP, where its enzymatic activities are required for viral RNA synthesis. However, the mechanistic processes behind the recruitment of NS5 from the cytoplasm to the RER membrane remain unclear. Here, we utilize high-resolution confocal microscopy and sucrose density gradient ultracentrifugation to investigate whether the association of NS5 with other NS proteins contributes to its membrane recruitment and retention. We demonstrate that NS1 or NS3 partially influences the NS5 association with the membrane. We further demonstrate that processed NS5 is predominantly in the cytoplasm and nucleus, indicating that the processing of NS5 from the viral polyprotein does not contribute to its membrane localization. These observations suggest that other host or viral factors, such as the enwrapment of NS5 by the RO, may also be necessary for the complete membrane retention of NS5. Therefore, studies on the inhibitors that disrupt the membrane localization of WNV NS5 are warranted for antiviral drug development.<\/jats:p>","DOI":"10.3390\/v16081303","type":"journal-article","created":{"date-parts":[[2024,8,16]],"date-time":"2024-08-16T04:29:57Z","timestamp":1723782597000},"page":"1303","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Membrane Retention of West Nile Virus NS5 Depends on NS1 or NS3 for Enzymatic Activity"],"prefix":"10.3390","volume":"16","author":[{"given":"Alanna C.","family":"Tseng","sequence":"first","affiliation":[{"name":"Department of Tropical Medicine, Medical Microbiology and Pharmacology, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, HI 96813, USA"},{"name":"Molecular Biosciences and Bioengineering Graduate Program, College of Tropical Agriculture and Human Resources, University of Hawaii at Manoa, Honolulu, HI 96822, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2044-2784","authenticated-orcid":false,"given":"Vivek R.","family":"Nerurkar","sequence":"additional","affiliation":[{"name":"Department of Tropical Medicine, Medical Microbiology and Pharmacology, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, HI 96813, USA"},{"name":"Molecular Biosciences and Bioengineering Graduate Program, College of Tropical Agriculture and Human Resources, University of Hawaii at Manoa, Honolulu, HI 96822, USA"},{"name":"Pacific Center for Emerging Infectious Diseases Research, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, HI 96813, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kabi R.","family":"Neupane","sequence":"additional","affiliation":[{"name":"Division of Math and Sciences, Leeward Community College, Pearl City, HI 96782, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helmut","family":"Kae","sequence":"additional","affiliation":[{"name":"Division of Math and Sciences, Leeward Community College, Pearl City, HI 96782, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pakieli H.","family":"Kaufusi","sequence":"additional","affiliation":[{"name":"Department of Tropical Medicine, Medical Microbiology and Pharmacology, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, HI 96813, USA"},{"name":"Molecular Biosciences and Bioengineering Graduate Program, College of Tropical Agriculture and Human Resources, University of Hawaii at Manoa, Honolulu, HI 96822, USA"},{"name":"Pacific Center for Emerging Infectious Diseases Research, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, HI 96813, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,8,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1807","DOI":"10.1056\/NEJM200106143442401","article-title":"The outbreak of West Nile virus infection in the New York City area in 1999","volume":"344","author":"Nash","year":"2001","journal-title":"N. Engl. J. Med."},{"key":"ref_2","unstructured":"U.S. Centers for Disease Control and Prevention (CDC) (2013). West Nile virus and other arboviral diseases\u2013United States, 2012. 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