quantitative multicolor super-resolution microscopy reveals tetherin hiv-1 interaction定量多色超分辨率显微镜揭示tetherin hiv - 1相互作用.pdfVIP

Quantitative Multicolor Super-Resolution Microscopy Reveals Tetherin HIV-1 Interaction 1 2 1,3 3 2 2 Martin Lehmann , Susana Rocha , Bastien Mangeat , Fabien Blanchet , Hiroshi Uji-i , Johan Hofkens , Vincent Piguet1,3* 1 Departments of Microbiology and Molecular Medicine, Dermatology and Venereology, University Hospital and Medical School of Geneva, Geneva, Switzerland, 2 Laboratory for Photochemistry and Spectroscopy, Department of Chemistry, Katholieke Universiteit Leuven, Heverlee, Belgium, 3 Department of Dermatology and Wound Healing, Cardiff University School of Medicine and University Hospital of Wales, Cardiff, Wales, United Kingdom 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. Tetheri