Functional Characterization of the HuRCD83 mRNA Interaction 英文参考文献.docVIP

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Functional Characterization of the HuRCD83 mRNA Interaction 英文参考文献.doc

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Functional Characterization of the HuRCD83 mRNA Interaction 英文参考文献

FunctionalCharacterizationoftheHuR:CD83mRNA Interaction DorotheaPieper1,SusannSchirmer1,AlexanderT.Prechtel1,RalphH.Kehlenbach2,JoachimHauber1, JanChemnitz1* 1Department of Cell Biology and Virology, Heinrich Pette Institute - Leibniz Institute for Experimental Virology, Hamburg, Germany, 2Zentrum fu¨r Biochemie und MolekulareZellbiologie,Universita¨t Go¨ttingen,Go¨ttingen,Germany Abstract Maturationofdendriticcells(DC)ischaracterizedbyexpressionofCD83,asurfaceproteinthatappearstobenecessaryfor theeffectiveactivationofna?¨veT-cellsandT-helpercellsbyDC.LatelyitwasshownthatCD83expressionisregulatedon the posttranscriptional level by interaction of the shuttle protein HuR with a novel posttranscriptional regulatory RNA element(PRE),whichislocatedinthecodingregionoftheCD83transcript.Interestingly,thisinteractioncommitstheCD83 mRNAtoefficientnuclearexportviatheCRM1pathway.Todate,however,thestructuralbasisofthisinteraction,which potentiallyinvolvesthreedistinctRNArecognitionmotifs(RRM1–3)inHuRandacomplexthree-prongedRNAstem-loop elementinCD83mRNA,hasnotbeeninvestigatedindetail.Inthepresentworkweanalyzedthisinteractioninvitroandin vivousingvariousHuR-andCD83mRNAmutants.Weareabletodemonstratethatboth,RRM1andRRM2arecrucialfor binding, whereas RRM3 as well as the HuR hinge region contributed only marginally to this protein:RNA interaction. Furthermore,mutationofuridinerichpatcheswithinthePREdidnotdisturbHuR:CD83mRNAcomplexformationwhile,in contrast, the deletion of specific PRE subfragments from the CD83 mRNA prevented HuR binding in vitro and in vivo. Interestingly,theobservedinhibitionofHuRbindingtoCD83mRNAdoesnotleadtoanucleartrappingofthetranscript but rather redirected this transcript from the CRM1- towards the NXF1/TAP-specific nuclear export pathway. Thus, the presenceofafunctionalPREpermitsnucleocytoplasmictraffickingoftheCD83transcriptviatheCRM1pathway. Citation:PieperD,SchirmerS,PrechtelAT,KehlenbachRH,HauberJ,etal.(2011)FunctionalCharacterizationoftheHuR:CD83mRN