title: Quantitative multicolor super-resolution microscopy reveals tetherin HIV-1 interaction.
abstract: Virus assembly and interaction with host-cell proteins occur at length scales below the diffraction limit of visible light. Novel super-resolution microscopy techniques achieve nanometer resolution of fluorescently labeled molecules. The cellular restriction factor tetherin (also known as CD317, BST-2 or HM1.24) inhibits the release of human immunodeficiency virus 1 (HIV-1) through direct incorporation into viral membranes and is counteracted by the HIV-1 protein Vpu. For super-resolution analysis of HIV-1 and tetherin interactions, we established fluorescence labeling of HIV-1 proteins and tetherin that preserved HIV-1 particle formation and Vpu-dependent restriction, respectively. Multicolor super-resolution microscopy revealed important structural features of individual HIV-1 virions, virus assembly sites and their interaction with tetherin at the plasma membrane. Tetherin localization to micro-domains was dependent on both tetherin membrane anchors. Tetherin clusters containing on average 4 to 7 tetherin dimers were visualized at HIV-1 assembly sites. Combined biochemical and super-resolution analysis revealed that extended tetherin dimers incorporate both N-termini into assembling virus particles and restrict HIV-1 release. Neither tetherin domains nor HIV-1 assembly sites showed enrichment of the raft marker GM1. Together, our super-resolution microscopy analysis of HIV-1 interactions with tetherin provides new insights into the mechanism of tetherin-mediated HIV-1 restriction and paves the way for future studies of virus-host interactions.
authors.affiliation: Department of Microbiology and Molecular Medicine, University Hospital and Medical School of Geneva, Geneva, Switzerland.
authors.forename: Martin
authors.initials: M
authors.lastname: Lehmann
authors.forename: Susana
authors.initials: S
authors.lastname: Rocha
authors.forename: Bastien
authors.initials: B
authors.lastname: Mangeat
authors.forename: Fabien
authors.initials: F
authors.lastname: Blanchet
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Uji-I
authors.forename: Johan
authors.initials: J
authors.lastname: Hofkens
authors.forename: Vincent
authors.initials: V
authors.lastname: Piguet
mesh_terms: D015703:Antigens, CD / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism*; D003116:Color; D005455:Fluorescent Antibody Technique / Q000379:methods; D058851:GPI-Linked Proteins / Q000737:chemistry / Q000378:metabolism; D015497:HIV-1 / Q000378:metabolism*; D054298:Human Immunodeficiency Virus Proteins / Q000378:metabolism*; D006801:Humans; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission / Q000379:methods; D014162:Transfection; D014771:Virion / Q000276:immunology / Q000378:metabolism*; D000074010:Bone Marrow Stromal Antigen 2
chemical_list: D015703:Antigens, CD; D058851:GPI-Linked Proteins; D054298:Human Immunodeficiency Virus Proteins; C094572:BST2 protein, human; D000074010:Bone Marrow Stromal Antigen 2