Hepatitis C Virus Translation Preferentially Depends on Active RNA Replication 英文参考文献.docVIP

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Hepatitis C Virus Translation Preferentially Depends on Active RNA Replication 英文参考文献.doc

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Hepatitis C Virus Translation Preferentially Depends on Active RNA Replication 英文参考文献

HepatitisCVirusTranslationPreferentiallyDependson ActiveRNAReplication HeleneMinyiLiu1,HidekiAizaki1,KeigoMachida1,J.-H.JamesOu1,MichaelM.C.Lai1,2* 1DepartmentofMolecularMicrobiologyandImmunology,KeckSchoolofMedicine,UniversityofSouthernCalifornia,LosAngeles,California,UnitedStatesofAmerica, 2InstituteofMolecularBiology,AcademiaSinica,Nankang,Taipei,Taiwan Abstract Hepatitis C virus (HCV) RNA initiates its replication on a detergent-resistant membrane structure derived from the endoplasmicreticulum(ER)intheHCVrepliconcells.Byperformingapulse-chasestudyofBrU-labeledHCVRNA,wefound that the newly-synthesized HCV RNA traveled along the anterograde-membrane traffic and moved away from the ER. Presumably,theRNAmovedtothesiteoftranslationorvirionassemblyinthelaterstepsofvirallifecycle.Inthisstudy,we furtheraddressedhowHCVRNAtranslationwasregulatedbyHCVRNAtrafficking.WhenthemovementofHCVRNAfrom thesiteofRNAsynthesistotheGolgicomplexwasblockedbynocodazole,aninhibitorofER-Golgitransport,HCVprotein translation was surprisingly enhanced, suggesting that the translation of viral proteins occurred near the site of RNA synthesis.WealsofoundthatthetranslationofHCVproteinswasdependentonactiveRNAsynthesis:inhibitionofviralRNA synthesisbyanNS5BinhibitorresultedindecreasedHCVviralproteinsynthesisevenwhenthetotalamountofintracellular HCVRNAremainedunchanged.Furthermore,thetranslationactivityofthereplication-defectiveHCVrepliconsorviralRNA withanNS5BmutationwasgreatlyreducedascomparedtothatofthecorrespondingwildtypeRNA.Byperforminglivecell labeling of newly synthesized HCV RNA and proteins, we further showed that the newly synthesized HCV proteins colocalizedwiththenewlysynthesizedviralRNA,suggestingthatHCVRNAreplicationandproteintranslationtakeplaceat ornearthesamesite.OurfindingstogetherindicatethatthetranslationofHCVRNAiscoupledtoRNAreplicationandthat thebothprocessesmayoccuratthesamesubcellularmembranecompartments,whichwetermthereplicasome. Citation:LiuHM,AizakiH,MachidaK,OuJ-HJ,La