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dc.contributor.authorPassos, Vânia
dc.contributor.authorZillinger, Thomas
dc.contributor.authorCasartelli, Nicoletta
dc.contributor.authorWachs, Amelie S
dc.contributor.authorXu, Shuting
dc.contributor.authorMalassa, Angelina
dc.contributor.authorSteppich, Katja
dc.contributor.authorSchilling, Hildegard
dc.contributor.authorFranz, Sergej
dc.contributor.authorTodt, Daniel
dc.contributor.authorSteinmann, Eike
dc.contributor.authorSutter, Kathrin
dc.contributor.authorDittmer, Ulf
dc.contributor.authorBohne, Jens
dc.contributor.authorSchwartz, Olivier
dc.contributor.authorBarchet, Winfried
dc.contributor.authorGoffinet, Christine
dc.date.accessioned2019-11-14T10:41:02Z
dc.date.available2019-11-14T10:41:02Z
dc.date.issued2019-10-09
dc.identifier.citationJ Virol. 2019 Oct 9. pii: JVI.01221-19. doi: 10.1128/JVI.01221-19en_US
dc.identifier.issn1098-5514
dc.identifier.pmid31597782
dc.identifier.doi10.1128/JVI.01221-19
dc.identifier.urihttp://hdl.handle.net/10033/622015
dc.description.abstractWhen expressed in virus-producing cells, the cellular multipass-transmembrane protein SERINC5 reduces the infectivity of HIV-1 particles and is counteracted by HIV-1 Nef. Due to the unavailability of an antibody of sufficient specificity and sensitivity, investigation of SERINC5 protein expression and subcellular localization has been limited to heterologously expressed SERINC5. We generated, via CRISPR/Cas9-assisted gene editing, Jurkat T-cell clones expressing endogenous SERINC5 bearing an extracellularly exposed HA epitope (Jurkat SERINC5(iHA-knock-in) T-cells). This modification enabled quantification of endogenous SERINC5 protein levels and demonstrated a predominant localization in lipid rafts. IFN-α treatment enhanced cell surface levels of SERINC5 in a Ruxolitinib-sensitive manner in the absence of modulation of mRNA and protein quantities. Parental and SERINC5(iHA-knock-in) T-cells shared the ability to produce infectious wildtype HIV-1, but not HIV-1 Δnef SERINC5-imposed reduction of infectivity involved a modest reduction of virus fusogenicity. Association of endogenous SERINC5 protein with HIV-1 Δnef virions was consistently detectable as a 35 kDa species, as opposed to heterologous SERINC5 that presented as 51 kDa species. Nef-mediated functional counteraction did not correlate with virion exclusion of SERINC5, arguing for the existence of additional counteractive mechanisms of Nef that act on virus-associated SERINC5. In HIV-1-infected cells, Nef triggered internalization of SERINC5 in the absence of detectable changes of steady-state protein levels. These findings establish new properties of endogenous SERINC5 expression and subcellular localization, challenge existing concepts of HIV-1 Nef-mediated antagonism of SERINC5 and uncover an unprecedented role of IFN-α in modulating SERINC5 through accumulation at the cell surface.IMPORTANCE SERINC5 is the long-searched antiviral factor that is counteracted by the HIV-1 accessory gene product Nef. Here, we engineered, via CRISPR/Cas9 technology, T-cell lines that express endogenous SERINC5 alleles tagged with a knocked-in HA epitope. This genetic modification enabled us to study basic properties of endogenous SERINC5 and to verify proposed mechanisms of HIV-1 Nef-mediated counteraction of SERINC5. Using this unique resource, we identified the susceptibility of endogenous SERINC5 protein to posttranslational modulation by type I IFNs and suggest uncoupling of Nef-mediated functional antagonism from SERINC5 exclusion from virions.en_US
dc.language.isoenen_US
dc.publisherAmerican Society of Microbiologyen_US
dc.rightsAttribution-NonCommercial-ShareAlike 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.titleCharacterization of Endogenous SERINC5 Protein as anti-HIV-1 Factor.en_US
dc.typeArticleen_US
dc.contributor.departmentTWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.en_US
dc.identifier.journalJournal of Virologyen_US
refterms.dateFOA2019-11-14T10:41:02Z
dc.source.journaltitleJournal of virology


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Attribution-NonCommercial-ShareAlike 4.0 International
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