Functional Specialization of the Plant miR396 Regulatory Network through Distinct MicroRNA–Target Interactions 英文参考文献.docVIP

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Functional Specialization of the Plant miR396 Regulatory Network through Distinct MicroRNA–Target Interactions 英文参考文献.doc

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Functional Specialization of the Plant miR396 Regulatory Network through Distinct MicroRNA–Target Interactions 英文参考文献

FunctionalSpecializationofthePlantmiR396Regulatory NetworkthroughDistinctMicroRNA–TargetInteractions JuanM.Debernardi.,RamiroE.Rodriguez.,MartinA.Mecchia,JavierF.Palatnik* InstitutodeBiolog?′aMolecularyCelulardeRosario(IBR),ConsejoNacionaldeInvestigacionesCient?′ficasyTe′cnicas(CONICET)andFacultaddeCienciasBioqu?′micasy Farmace′uticas,UniversidadNacionaldeRosario,Rosario,Argentina Abstract MicroRNAs(miRNAs)are,21ntsmallRNAsthatregulategeneexpressioninanimalsandplants.Theycanbegroupedinto families comprising different genes encoding similar or identical mature miRNAs. Several miRNA families are deeply conservedinplantlineagesandregulatekeyaspectsofplantdevelopment,hormonesignaling,andstressresponse.The ancient miRNA miR396 regulates conserved targets belonging to the GROWTH-REGULATING FACTOR (GRF) family of transcriptionfactors,whichareknowntocontrolcellproliferationinArabidopsisleaves.Inthiswork,wecharacterizedthe regulation of an additional target for miR396, the transcription factor bHLH74, that is necessary for Arabidopsis normal development.bHLH74homologswithamiR396targetsitecouldonlybedetectedinthesisterfamiliesBrassicaceaeand Cleomaceae. Still, bHLH74 repression by miR396 is required for margin and vein pattern formation of Arabidopsis leaves. MiR396contributestothespatio-temporalregulationofGRFandbHLH74expressionduringleafdevelopment.Furthermore, asurveyofmiR396sequencesindifferentspeciesshowedvariationsinthe59portionofthemiRNA,aregionknowntobe important for miRNA activity. Analysis of different miR396 variants in Arabidopsis thaliana revealed that they have an enhancedactivitytowardGRFtranscriptionfactors.TheinteractionbetweentheGRFtargetsiteandmiR396hasabulge betweenpositions7and8ofthemiRNA.OurdataindicatethatsuchbulgemodulatesthestrengthofthemiR396-mediated repressionandthatthismodulationisessentialtoshapetheprecisespatio-temporalpatternofGRF2expression.Theresults showthatancientmiRNAscanregulateconservedtargetswithvariedefficiencyindifferentspecies,an