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While its antiviral function is well established in retroviral systems, its role in SARS-CoV-2 egress remains unclear. Here, we used a virus-like particle (VLP) system composed of SARS-CoV-2 structural proteins M, E, and N to investigate the impact of BST-2 on viral particle release. BST-2 significantly inhibited VLP release in HEK293T and Calu-3 lung epithelial cells. Confocal microscopy revealed that BST-2 colocalizes with viral structural proteins at the endoplasmic reticulum-Golgi intermediate compartment (ERGIC), the main site of coronavirus assembly. We next evaluated the roles of the SARS-CoV-2 accessory proteins ORF3a and ORF7a in overcoming this restriction. ORF3a localized to endolysosomal compartments and promoted VLP release through a BST-2-independent mechanism, without altering BST-2 expression or localization. In contrast, ORF7a colocalized with both BST-2 and ERGIC markers and restored VLP release by promoting BST-2 degradation. Notably, ORF7a also relieved BST-2-mediated restriction of HIV-1 VLP release, suggesting a conserved antagonistic function. These findings identify BST-2 as an intracellular inhibitor of SARS-CoV-2 particle release and establish ORF7a as viral accessory antagonist that neutralizes this host defense.<\/jats:p>","DOI":"10.1101\/2025.07.07.663519","type":"posted-content","created":{"date-parts":[[2025,7,11]],"date-time":"2025-07-11T10:20:10Z","timestamp":1752229210000},"source":"Crossref","is-referenced-by-count":2,"title":["BST-2 inhibits SARS-CoV-2 egress at intracellular membranes and is neutralized by ORF7a"],"prefix":"10.64898","author":[{"given":"Adam","family":"Smith","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinhong","family":"Dong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025071103200509000_2025.07.07.663519v1.1","doi-asserted-by":"publisher","DOI":"10.1101\/cshperspect.a006940"},{"key":"2025071103200509000_2025.07.07.663519v1.2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1011485107"},{"key":"2025071103200509000_2025.07.07.663519v1.3","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2009.08.039"},{"issue":"4","key":"2025071103200509000_2025.07.07.663519v1.4","doi-asserted-by":"crossref","first-page":"321","DOI":"10.2174\/157016212800792522","article-title":"Intracellular logistics of BST-2\/tetherin","volume":"10","year":"2012","journal-title":"Curr HIV Res"},{"key":"2025071103200509000_2025.07.07.663519v1.5","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02211-08"},{"key":"2025071103200509000_2025.07.07.663519v1.6","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0811014106"},{"key":"2025071103200509000_2025.07.07.663519v1.7","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00103-10"},{"key":"2025071103200509000_2025.07.07.663519v1.8","doi-asserted-by":"publisher","DOI":"10.1089\/aid.2009.0253"},{"key":"2025071103200509000_2025.07.07.663519v1.9","doi-asserted-by":"publisher","DOI":"10.1042\/BCJ20160861"},{"key":"2025071103200509000_2025.07.07.663519v1.10","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02274-15"},{"key":"2025071103200509000_2025.07.07.663519v1.11","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1016\/j.virol.2013.11.030","article-title":"BST2\/CD317 counteracts human coronavirus 229E productive infection by tethering virions at the cell surface","volume":"449","year":"2014","journal-title":"Virology"},{"issue":"10","key":"2025071103200509000_2025.07.07.663519v1.12","doi-asserted-by":"crossref","first-page":"1737","DOI":"10.1080\/15548627.2019.1707487","article-title":"BST2 suppresses porcine epidemic diarrhea virus replication by targeting and degrading virus nucleocapsid protein with selective autophagy","volume":"16","year":"2020","journal-title":"Autophagy"},{"key":"2025071103200509000_2025.07.07.663519v1.13","doi-asserted-by":"crossref","first-page":"110607","DOI":"10.1016\/j.virol.2025.110607","article-title":"SARS-CoV-2 host-pathogen interactome: insights into more players during pathogenesis","volume":"610","year":"2025","journal-title":"Virology"},{"key":"2025071103200509000_2025.07.07.663519v1.14","doi-asserted-by":"crossref","unstructured":"Katiyar, H. , et al., SARS-CoV-2 Assembly: Gaining Infectivity and Beyond. 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