{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T20:14:47Z","timestamp":1790626487383,"version":"4.1.0"},"reference-count":47,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2020,4,18]],"date-time":"2020-04-18T00:00:00Z","timestamp":1587168000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Bone Marrow Stromal Cell Antigen 2 (BST-2)\/tetherin inhibits the release of numerous enveloped viruses by physically tethering nascent particles to infected cells during the process of viral budding from the cell surface. Tetherin also restricts human immunodeficiency virus (HIV), and pandemic main (M) group HIV type 1s (HIV-1s) are thought to rely exclusively on their Vpu proteins to overcome tetherin-mediated restriction of virus release. However, at least one M group HIV-1 strain, the macrophage-tropic primary AD8 isolate, is unable to express Vpu due to a mutation in its translation initiation codon. Here, using primary monocyte-derived macrophages (MDMs), we show that AD8 Nef protein can compensate for the absence of Vpu and restore virus release to wild type levels. We demonstrate that HIV-1 AD8 Nef reduces endogenous cell surface tetherin levels, physically separating it from the site of viral budding, thus preventing HIV retention. Mechanistically, AD8 Nef enhances internalisation of the long isoform of human tetherin, leading to perinuclear accumulation of the restriction factor. Finally, we show that Nef proteins from other HIV strains also display varying degrees of tetherin antagonism. Overall, we show that M group HIV-1s can use an accessory protein other than Vpu to antagonise human tetherin.<\/jats:p>","DOI":"10.3390\/v12040459","type":"journal-article","created":{"date-parts":[[2020,4,21]],"date-time":"2020-04-21T04:49:38Z","timestamp":1587444578000},"page":"459","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["The Nef Protein of the Macrophage Tropic HIV-1 Strain AD8 Counteracts Human BST-2\/Tetherin"],"prefix":"10.3390","volume":"12","author":[{"given":"Sebastian","family":"Giese","sequence":"first","affiliation":[{"name":"MRC Laboratory for Molecular Cell Biology, University College London, London WC1E 6BT, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2887-3933","authenticated-orcid":false,"given":"Scott P.","family":"Lawrence","sequence":"additional","affiliation":[{"name":"MRC Laboratory for Molecular Cell Biology, University College London, London WC1E 6BT, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2462-9925","authenticated-orcid":false,"given":"Michela","family":"Mazzon","sequence":"additional","affiliation":[{"name":"MRC Laboratory for Molecular Cell Biology, University College London, London WC1E 6BT, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bernadien M.","family":"Nijmeijer","sequence":"additional","affiliation":[{"name":"MRC Laboratory for Molecular Cell Biology, University College London, London WC1E 6BT, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0213-3259","authenticated-orcid":false,"given":"Mark","family":"Marsh","sequence":"additional","affiliation":[{"name":"MRC Laboratory for Molecular Cell Biology, University College London, London WC1E 6BT, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,4,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1016\/j.chom.2009.11.002","article-title":"A tail of Tetherin: How pandemic HIV-1 conquered the world","volume":"6","author":"Gupta","year":"2009","journal-title":"Cell Host Microbe"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1016\/j.chom.2009.10.004","article-title":"Tetherin-driven adaptation of Vpu and Nef function and the evolution of pandemic and nonpandemic HIV-1 strains","volume":"6","author":"Sauter","year":"2009","journal-title":"Cell Host Microbe"},{"key":"ref_3","first-page":"67","article-title":"The antiviral activities of tetherin","volume":"371","author":"Neil","year":"2013","journal-title":"Curr. Top. Microbiol. Immunol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Venkatesh, S., and Bieniasz, P.D. (2013). Mechanism of HIV-1 virion entrapment by tetherin. PLoS Pathog., 9.","DOI":"10.1371\/journal.ppat.1003483"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Cocka, L.J., and Bates, P. (2012). Identification of alternatively translated Tetherin isoforms with differing antiviral and signaling activities. PLoS Pathog., 8.","DOI":"10.1371\/journal.ppat.1002931"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1016\/j.chom.2012.10.007","article-title":"Innate sensing of HIV-1 assembly by Tetherin induces NFkappaB-dependent proinflammatory responses","volume":"12","author":"Galao","year":"2012","journal-title":"Cell Host Microbe"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1038\/nature06553","article-title":"Tetherin inhibits retrovirus release and is antagonized by HIV-1 Vpu","volume":"451","author":"Neil","year":"2008","journal-title":"Nature"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/j.chom.2008.03.001","article-title":"The interferon-induced protein BST-2 restricts HIV-1 release and is downregulated from the cell surface by the viral Vpu protein","volume":"3","author":"Goff","year":"2008","journal-title":"Cell Host Microbe"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Gupta, R.K., Hue, S., Schaller, T., Verschoor, E., Pillay, D., and Towers, G.J. (2009). Mutation of a single residue renders human tetherin resistant to HIV-1 Vpu-mediated depletion. PLoS Pathog., 5.","DOI":"10.1371\/journal.ppat.1000443"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"McNatt, M.W., Zang, T., Hatziioannou, T., Bartlett, M., Fofana, I.B., Johnson, W.E., Neil, S.J., and Bieniasz, P.D. (2009). Species-specific activity of HIV-1 Vpu and positive selection of tetherin transmembrane domain variants. PLoS Pathog., 5.","DOI":"10.1371\/journal.ppat.1000300"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Freed, E.O. (2013). Cellular Trafficking Mechanisms in the Assembly and Release of HIV. Advances in HIV-1 Assembly and Release, Springer.","DOI":"10.1007\/978-1-4614-7729-7"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"7931","DOI":"10.1128\/JVI.00242-09","article-title":"Vpu directs the degradation of the human immunodeficiency virus restriction factor BST-2\/Tetherin via a {beta}TrCP-dependent mechanism","volume":"83","author":"Douglas","year":"2009","journal-title":"J. Virol."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Janvier, K., Pelchen-Matthews, A., Renaud, J.B., Caillet, M., Marsh, M., and Berlioz-Torrent, C. (2011). The ESCRT-0 component HRS is required for HIV-1 Vpu-mediated BST-2\/tetherin down-regulation. PLoS Pathog., 7.","DOI":"10.1371\/journal.ppat.1001265"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Mangeat, B., Gers-Huber, G., Lehmann, M., Zufferey, M., Luban, J., and Piguet, V. (2009). HIV-1 Vpu neutralizes the antiviral factor Tetherin\/BST-2 by binding it and directing its beta-TrCP2-dependent degradation. PLoS Pathog., 5.","DOI":"10.1371\/journal.ppat.1000574"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"McNatt, M.W., Zang, T., and Bieniasz, P.D. (2013). Vpu binds directly to tetherin and displaces it from nascent virions. PLoS Pathog., 9.","DOI":"10.1371\/journal.ppat.1003299"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Mitchell, R.S., Katsura, C., Skasko, M.A., Fitzpatrick, K., Lau, D., Ruiz, A., Stephens, E.B., Margottin-Goguet, F., Benarous, R., and Guatelli, J.C. (2009). Vpu antagonizes BST-2-mediated restriction of HIV-1 release via beta-TrCP and endo-lysosomal trafficking. PLoS Pathog., 5.","DOI":"10.1371\/journal.ppat.1000450"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"11966","DOI":"10.1128\/JVI.01515-09","article-title":"Antagonism to and intracellular sequestration of human tetherin by the human immunodeficiency virus type 2 envelope glycoprotein","volume":"83","author":"Neil","year":"2009","journal-title":"J. Virol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"20889","DOI":"10.1073\/pnas.0907075106","article-title":"Simian immunodeficiency virus envelope glycoprotein counteracts tetherin\/BST-2\/CD317 by intracellular sequestration","volume":"106","author":"Gupta","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"639","DOI":"10.1016\/j.chom.2014.10.002","article-title":"Nef proteins of epidemic HIV-1 group O strains antagonize human tetherin","volume":"16","author":"Kluge","year":"2014","journal-title":"Cell Host Microbe"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"659","DOI":"10.1016\/j.cell.2018.07.004","article-title":"HIV-1 Nefs Are Cargo-Sensitive AP-1 Trimerization Switches in Tetherin Downregulation","volume":"174","author":"Morris","year":"2018","journal-title":"Cell"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/j.chom.2009.05.008","article-title":"Nef proteins from simian immunodeficiency viruses are tetherin antagonists","volume":"6","author":"Zhang","year":"2009","journal-title":"Cell Host Microbe"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1016\/j.chom.2010.12.005","article-title":"Compensatory changes in the cytoplasmic tail of gp41 confer resistance to tetherin\/BST-2 in a pathogenic nef-deleted SIV","volume":"9","author":"Jia","year":"2011","journal-title":"Cell Host Microbe"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Zhang, F., Landford, W.N., Ng, M., McNatt, M.W., Bieniasz, P.D., and Hatziioannou, T. (2011). SIV Nef proteins recruit the AP-2 complex to antagonize Tetherin and facilitate virion release. PLoS Pathog., 7.","DOI":"10.1371\/journal.ppat.1002039"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Jia, B., Serra-Moreno, R., Neidermyer, W., Rahmberg, A., Mackey, J., Fofana, I.B., Johnson, W.E., Westmoreland, S., and Evans, D.T. (2009). Species-specific activity of SIV Nef and HIV-1 Vpu in overcoming restriction by tetherin\/BST2. PLoS Pathog., 5.","DOI":"10.1371\/journal.ppat.1000429"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"887","DOI":"10.1128\/JVI.73.2.887-896.1999","article-title":"Regulation of virus release by the macrophage-tropic human immunodeficiency virus type 1 AD8 isolate is redundant and can be controlled by either Vpu or Env","volume":"73","author":"Schubert","year":"1999","journal-title":"J. Virol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2906","DOI":"10.1128\/jvi.68.5.2906-2914.1994","article-title":"Optimal infectivity in vitro of human immunodeficiency virus type 1 requires an intact nef gene","volume":"68","author":"Chowers","year":"1994","journal-title":"J. Virol."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Giese, S., and Marsh, M. (2014). Tetherin can restrict cell-free and cell-cell transmission of HIV from primary macrophages to T cells. PLoS Pathog., 10.","DOI":"10.1371\/journal.ppat.1004189"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"5845","DOI":"10.1128\/JVI.03818-13","article-title":"Differential sensitivities of tetherin isoforms to counteraction by primate lentiviruses","volume":"88","author":"Weinelt","year":"2014","journal-title":"J. Virol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1083\/jcb.200304008","article-title":"Infectious HIV-1 assembles in late endosomes in primary macrophages","volume":"162","author":"Kramer","year":"2003","journal-title":"J. Cell Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"7699","DOI":"10.1128\/jvi.69.12.7699-7711.1995","article-title":"Augmentation of virus secretion by the human immunodeficiency virus type 1 Vpu protein is cell type independent and occurs in cultured human primary macrophages and lymphocytes","volume":"69","author":"Schubert","year":"1995","journal-title":"J. Virol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1089\/aid.1996.12.191","article-title":"Construction and characterization of a stable full-length macrophage-tropic HIV type 1 molecular clone that directs the production of high titers of progeny virions","volume":"12","author":"Theodore","year":"1996","journal-title":"AIDS Res. Hum. Retrovir."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"212","DOI":"10.1038\/nature15399","article-title":"HIV-1 Nef promotes infection by excluding SERINC5 from virion incorporation","volume":"526","author":"Rosa","year":"2015","journal-title":"Nature"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1038\/nature15400","article-title":"SERINC3 and SERINC5 restrict HIV-1 infectivity and are counteracted by Nef","volume":"526","author":"Usami","year":"2015","journal-title":"Nature"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Zutz, A., Scholz, C., Schneider, S., Pierini, V., Munchhoff, M., Sutter, K., Wittmann, G., Dittmer, U., Draenert, R., and Bogner, J.R. (2020). SERINC5 Is an Unconventional HIV Restriction Factor That Is Upregulated during Myeloid Cell Differentiation. J. Innate Immun., 1\u201311.","DOI":"10.1159\/000504888"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1016\/0042-6822(92)90293-X","article-title":"Effect of myristoylation on p27 nef subcellular distribution and suppression of HIV-LTR transcription","volume":"187","author":"Yu","year":"1992","journal-title":"Virology"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2780","DOI":"10.1099\/vir.0.83120-0","article-title":"Effects of vpu start-codon mutations on human immunodeficiency virus type 1 replication in macrophages","volume":"88","author":"Richards","year":"2007","journal-title":"J. Gen. Virol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1128\/JVI.01795-10","article-title":"Serine-threonine ubiquitination mediates downregulation of BST-2\/tetherin and relief of restricted virion release by HIV-1 Vpu","volume":"85","author":"Tokarev","year":"2011","journal-title":"J. Virol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3850","DOI":"10.1242\/jcs.003343","article-title":"Clathrin-mediated endocytosis of a lipid-raft-associated protein is mediated through a dual tyrosine motif","volume":"120","author":"Rollason","year":"2007","journal-title":"J. Cell Sci."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Kueck, T., and Neil, S.J. (2012). A cytoplasmic tail determinant in HIV-1 Vpu mediates targeting of tetherin for endosomal degradation and counteracts interferon-induced restriction. PLoS Pathog., 8.","DOI":"10.1371\/journal.ppat.1002609"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"496","DOI":"10.2174\/157016211798842116","article-title":"One protein to rule them all: Modulation of cell surface receptors and molecules by HIV Nef","volume":"9","author":"Landi","year":"2011","journal-title":"Curr. HIV Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1016\/j.chom.2015.09.003","article-title":"Cell Surface Proteomic Map of HIV Infection Reveals Antagonism of Amino Acid Metabolism by Vpu and Nef","volume":"18","author":"Matheson","year":"2015","journal-title":"Cell Host Microbe"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"10701","DOI":"10.1128\/JVI.01465-16","article-title":"Tetherin Antagonism by HIV-1 Group M Nef Proteins","volume":"90","author":"Arias","year":"2016","journal-title":"J. Virol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"179","DOI":"10.20411\/pai.v2i2.204","article-title":"HIV Infection of Macrophages: Implications for Pathogenesis and Cure","volume":"2","author":"Clayton","year":"2017","journal-title":"Pathog. Immun."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1435","DOI":"10.3389\/fimmu.2019.01435","article-title":"The HIV Reservoir in Monocytes and Macrophages","volume":"10","author":"Wong","year":"2019","journal-title":"Front. Immunol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2899","DOI":"10.1099\/0022-1317-81-12-2899","article-title":"T-cell line adaptation of human immunodeficiency virus type 1 strain SF162: Effects on envelope, vpu and macrophage-tropism","volume":"81","author":"Dejucq","year":"2000","journal-title":"J. Gen. Virol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"3877","DOI":"10.1128\/JVI.02725-06","article-title":"Downregulation of CD4 by human immunodeficiency virus type 1 Nef is dependent on clathrin and involves direct interaction of Nef with the AP2 clathrin adaptor","volume":"81","author":"Chaudhuri","year":"2007","journal-title":"J. Virol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1166","DOI":"10.1128\/JVI.01874-07","article-title":"A diacidic motif in human immunodeficiency virus type 1 Nef is a novel determinant of binding to AP-2","volume":"82","author":"Lindwasser","year":"2008","journal-title":"J. Virol."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/12\/4\/459\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T13:45:06Z","timestamp":1760363106000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/12\/4\/459"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,4,18]]},"references-count":47,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2020,4]]}},"alternative-id":["v12040459"],"URL":"https:\/\/doi.org\/10.3390\/v12040459","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2020.02.07.938464","asserted-by":"object"}]},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,4,18]]}}}