Molecular Determinants and Dynamics of Hepatitis C Virus Secretion 英文参考文献.docVIP

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Molecular Determinants and Dynamics of Hepatitis C Virus Secretion 英文参考文献.doc

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Molecular Determinants and Dynamics of Hepatitis C Virus Secretion 英文参考文献

MolecularDeterminantsandDynamicsofHepatitisC VirusSecretion KellyE.Coller,NicholasS.Heaton,KristiL.Berger,JacobD.Cooper,JessicaL.Saunders,GlennRandall* DepartmentofMicrobiology,TheUniversityofChicago,Chicago,Illinois,UnitedStatesofAmerica Abstract The current model of hepatitis C virus (HCV) production involves the assembly of virions on or near the surface of lipid droplets,envelopmentattheERinassociationwithcomponentsofVLDLsynthesis,andegressviathesecretorypathway. However, the cellular requirements for and a mechanistic understanding of HCV secretion are incomplete at best. We combinedanRNAinterference(RNAi)analysisofhostfactorsforinfectiousHCVsecretionwiththedevelopmentoflivecell imaging of HCV core trafficking to gain a detailed understanding of HCV egress. RNAi studies identified multiple components of the secretory pathway, including ER to Golgi trafficking, lipid and protein kinases that regulate budding fromthetrans-Golginetwork(TGN),VAMP1vesiclesandadaptorproteins,andtherecyclingendosome.Ourresultssupport a model wherein HCV is infectious upon envelopment at the ER and exits the cell via the secretory pathway. We next constructedinfectiousHCVwithatetracysteine(TC)taginsertionincore(TC-core)tomonitorthedynamicsofHCVcore traffickinginassociationwithitscellularcofactors.Inordertoisolatecoreproteinmovementsassociatedwithinfectious HCVsecretion,onlytraffickingeventsthatrequiredtheessentialHCVassemblyfactorNS2werequantified.TC-coretraffics tothecellperipheryalongmicrotubulesandthismovementcanbeinhibitedbynocodazole.Sub-populationsofTC-core localize to the Golgi and co-traffic with components of the recycling endosome. Silencing of the recycling endosome componentRab11aresultsintheaccumulationofHCVcoreattheGolgi.Themajorityofdynamiccoretrafficsinassociation with apolipoprotein E (ApoE) and VAMP1 vesicles. This study identifies many new host cofactors of HCV egress, while presentingdynamicstudiesofHCVcoretraffickingininfectedcells. Citation: Coller KE, Heaton NS