An RNA Element at the 5′-End of the Poliovirus Genome Functions as a General Promoter for RNA Synthesis 英文参考文献.docVIP
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An RNA Element at the 5′-End of the Poliovirus Genome Functions as a General Promoter for RNA Synthesis 英文参考文献
AnRNAElementatthe59-EndofthePoliovirusGenome
FunctionsasaGeneralPromoterforRNASynthesis
DorotheeA.Vogt¤,RaulAndino*
DepartmentofMicrobiologyandImmunology,UniversityofCalifornia,SanFrancisco,California,UnitedStatesofAmerica
Abstract
RNA structures present throughout RNA virus genomes serve as scaffolds to organize multiple factors involved in the
initiation of RNA synthesis. Several of these RNA elements play multiple roles in the RNA replication pathway. An RNA
structureformedaroundthe59-endofthepoliovirusgenomicRNAhasbeenimplicatedintheinitiationofbothnegative-
and positive-strand RNA synthesis. Dissecting the roles of these multifunctional elements is usually hindered by the
interdependent nature of the viral replication processes and often pleiotropic effects of mutations. Here, we describe a
novelapproachtoexamineRNAelementswithmultipleroles.OurapproachreliesontheduplicationoftheRNAstructure
sothatonecopyisdedicatedtotheinitiationofnegative-strandRNAsynthesis,whiletheothermediatespositive-strand
synthesis.Thisallowsustostudythefunctionoftheelementinpromotingpositive-strandRNAsynthesis,independentlyof
its function in negative-strand initiation.Using this approach, we demonstrate that the entire 59-end RNA structure that
formsonthepositive-strandisrequiredforinitiationofnewpositive-strandRNAs.Alsorequiredtoinitiatepositive-strand
RNA synthesisare thebinding sitesfor theviralpolymerase precursor, 3CD,andthe host factor, PCBP. Furthermore, we
identifyspecificnucleotidesequenceswithin‘‘stema’’thatareessentialfortheinitiationofpositive-strandRNAsynthesis.
Thesefindingsprovidedirectevidenceforatrans-initiationmodel,inwhichbindingofproteinstointernalsequencesofa
pre-existing positive-strand RNA affects the synthesis of subsequent copies of that RNA, most likely by organizing
replicationfactorsaroundtheinitiationsite.
Citation:VogtDA,AndinoR(2010)AnRNAElementatthe59-EndofthePoliovirusGenomeFunctionsasaGeneralPromoterforRNASynthesis.PLoSPathog6(6):
e1000936.doi:10.1371
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