G Protein-Coupled Receptor Kinase 2 Promotes Flaviviridae Entry and Replication 英文参考文献.docVIP

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G Protein-Coupled Receptor Kinase 2 Promotes Flaviviridae Entry and Replication 英文参考文献.doc

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G Protein-Coupled Receptor Kinase 2 Promotes Flaviviridae Entry and Replication 英文参考文献

GProtein-CoupledReceptorKinase2Promotes FlaviviridaeEntryandReplication CarolineLeSommer1,2,NicholasJ.Barrows2,3,4,SheltonS.Bradrick1,2,JamesL.Pearson1,2,3 , MarianoA.Garcia-Blanco1,2,5,6 * 1DepartmentofMolecularGeneticsandMicrobiology,DukeUniversityMedicalCenter,Durham,NorthCarolina,UnitedStatesofAmerica,2CenterforRNABiology,Duke UniversityMedicalCenter,Durham,NorthCarolina,UnitedStatesofAmerica,3DukeRNAiScreeningFacility,DukeUniversityMedicalCenter,Durham,NorthCarolina, UnitedStatesofAmerica,4InstituteforGenomeSciencesandPolicy,DukeUniversity,Durham,NorthCarolina,UnitedStatesofAmerica,5DepartmentofMedicine,Duke UniversityMedicalCenter,Durham,NorthCarolina,UnitedStatesofAmerica,6PrograminEmergingInfectiousDiseases,Duke-NUSGraduateMedicalSchool,Singapore, RepublicofSingapore Abstract Flavivirusescauseawiderangeofseverediseasesrangingfromencephalitistohemorrhagicfever.Discoveryofhostfactors thatregulatethefateofflavivirusesininfectedcellscouldprovideinsightintothemolecularmechanismsofinfectionand thereforefacilitatethedevelopmentofanti-flaviviraldrugs.Weperformedgenome-scalesiRNAscreenstodiscoverhuman hostfactorsrequiredforyellowfevervirus(YFV)propagation.Usinga262siRNApoolscreeningformatandaduplicateof the screen, we identified a high confidence list of YFV host factors. To find commonalities between flaviviruses, these candidateswerecomparedtohostfactorspreviouslyidentifiedforWestNilevirus(WNV)anddenguevirus(DENV).This comparison highlighted a potential requirement for the G protein-coupled receptor kinase family, GRKs, for flaviviral infection.TheYFVhostcandidateGRK2(alsoknownasADRBK1)wasvalidatedbothinsiRNA-mediatedknockdownHuH-7 cellsandinGRK2/2mouseembryonicfibroblasts.Additionally,weshowedthatGRK2wasrequiredforefficientpropagation ofDENVandHepatitisCvirus(HCV)indicatingthatGRK2requirementisconservedthroughouttheFlaviviridae.Finally,we foundthatGRK2participatesinmultipledistinctstepsoftheflaviviruslifecyclebypromotingbothentryandRNAsynthesis. Together, our fi