Quantitative Multicolor Super-Resolution Microscopy Reveals Tetherin HIV-1 Interaction 英文参考文献.docVIP

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Quantitative Multicolor Super-Resolution Microscopy Reveals Tetherin HIV-1 Interaction 英文参考文献.doc

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Quantitative Multicolor Super-Resolution Microscopy Reveals Tetherin HIV-1 Interaction 英文参考文献

QuantitativeMulticolorSuper-ResolutionMicroscopy RevealsTetherinHIV-1Interaction MartinLehmann1,SusanaRocha2,BastienMangeat1,3,FabienBlanchet3,HiroshiUji-i2,JohanHofkens2, VincentPiguet1,3 * 1Departments of Microbiology and Molecular Medicine, Dermatology and Venereology, University Hospital and Medical School of Geneva, Geneva, Switzerland, 2Laboratory for Photochemistry and Spectroscopy, Department of Chemistry, Katholieke Universiteit Leuven, Heverlee, Belgium, 3Department of Dermatology and WoundHealing,CardiffUniversitySchoolofMedicineandUniversityHospitalofWales,Cardiff,Wales,UnitedKingdom Abstract Virusassemblyandinteractionwithhost-cellproteinsoccuratlengthscalesbelowthediffractionlimitofvisiblelight.Novel super-resolution microscopy techniques achieve nanometer resolution of fluorescently labeled molecules. The cellular restrictionfactortetherin(alsoknownasCD317,BST-2orHM1.24)inhibitsthereleaseofhumanimmunodeficiencyvirus1 (HIV-1) through direct incorporation into viral membranes and is counteracted by the HIV-1 protein Vpu. For super- resolutionanalysisofHIV-1andtetherininteractions,weestablishedfluorescencelabelingofHIV-1proteinsandtetherin thatpreservedHIV-1particleformationandVpu-dependentrestriction,respectively.Multicolorsuper-resolutionmicroscopy revealedimportantstructuralfeaturesofindividualHIV-1virions,virusassemblysitesandtheirinteractionwithtetherinat 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 assemblingvirusparticlesandrestrictHIV-1release.NeithertetherindomainsnorHIV-1assemblysitesshowedenrichment oftheraftmarkerGM1.Together,oursuper-resolutionmicroscopyanalysisofHIV-1interactionswithtetherinprovidesnew insights into the mechanism of tetherin-mediated HIV