Small RNA Deep Sequencing Reveals Role for Arabidopsis thaliana RNA-Dependent RNA Polymerases in Viral siRNA Biogenesis 英文参考文献.docVIP

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Small RNA Deep Sequencing Reveals Role for Arabidopsis thaliana RNA-Dependent RNA Polymerases in Viral siRNA Biogenesis 英文参考文献.doc

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Small RNA Deep Sequencing Reveals Role for Arabidopsis thaliana RNA-Dependent RNA Polymerases in Viral siRNA Biogenesis 英文参考文献

SmallRNADeepSequencingRevealsRolefor ArabidopsisthalianaRNA-DependentRNAPolymerases inViralsiRNABiogenesis XiaopengQi1¤,ForrestShengBao2,ZhixinXie1* 1DepartmentofBiologicalSciences,TexasTechUniversity,Lubbock,Texas,UnitedStatesofAmerica,2DepartmentofComputerScience,TexasTechUniversity,Lubbock, Texas,UnitedStatesofAmerica Abstract RNAsilencingfunctionsasanimportantantiviraldefensemechanisminabroadrangeofeukaryotes.Inplants,biogenesis ofseveralclassesofendogenoussmallinterferingRNAs(siRNAs)requiresRNA-dependentRNAPolymerase(RDR)activities. MembersoftheRDRfamilyproteins,includingRDR1andRDR6,havealsobeenimplicatedinantiviraldefense,althougha directroleforRDRsinviralsiRNAbiogenesishasyettobedemonstrated.Usingacrucifer-infectingstrainofTobaccoMosaic Virus(TMV-Cg)andArabidopsisthalianaasamodelsystem,weanalyzedtheviralsmallRNAprofileinwild-typeplantsas wellasrdrmutantsbyapplyingsmallRNAdeepsequencingtechnology.Over100,000TMV-Cg-specificsmallRNAreads, mostlyof21-(78.4%)and22-nucleotide(12.9%)insizeandoriginatingpredominately(79.9%)fromthegenomicsenseRNA strand,werecapturedatanearlyinfectionstage,yieldingthefirsthigh-resolutionsmallRNAmapforaplantvirus.TheTMV- Cggenomeharboredmultiple,highlyreproduciblesmallRNA-generatinghotspotsthatcorrespondedtoregionswithno apparentlocalhairpin-formingcapacity.Significantly,boththerdr1andrdr6mutantsexhibitedgloballyreducedlevelsof viralsmallRNAproductionaswellasreducedstrandbiasinviralsmallRNApopulation,revealinganimportantroleforthese host RDRs in viral siRNA biogenesis. In addition, an informatics analysis showed that a large set of host genes could be potentiallytargetedbyTMV-Cg-derivedsiRNAsforposttranscriptionalsilencing.Twoofsuchpredictedhosttargets,which encodeacleavageandpolyadenylationspecificityfactor(CPSF30)andanunknownproteinsimilartotranslocon-associated proteinalpha(TRAPa),respectively,yieldedapositiveresultincleavagevalidationby59RACEassays.Ourdataraisedthe interestingpossibilityforviralsiRNA-mediatedvirus-hostinteractionsth