SAF-A Forms a Complex with BRG1 and Both Components Are Required for RNA Polymerase II Mediated Transcription 英文参考文献.docVIP

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SAF-A Forms a Complex with BRG1 and Both Components Are Required for RNA Polymerase II Mediated Transcription 英文参考文献.doc

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SAF-A Forms a Complex with BRG1 and Both Components Are Required for RNA Polymerase II Mediated Transcription 英文参考文献

SAF-AFormsaComplexwithBRG1andBoth ComponentsAreRequiredforRNAPolymeraseII MediatedTranscription DzenetaVizlin-Hodzic1,RikardRunnberg1,JessicaRyme1,StinaSimonsson2*,TomasSimonsson1* 1DepartmentofMedicalBiochemistryandCellBiology,InstituteofBiomedicine,UniversityofGothenburg,Gothenburg,Sweden,2DepartmentofClinicalChemistryand TransfusionMedicine,InstituteofBiomedicine,SahlgrenskaUniversityHospital,Gothenburg,Sweden Abstract Background: Scaffold attachment factor A (SAF-A) participates in the regulation of gene expression by organizing chromatinintotranscriptionallyactivedomainsandbyinteractingdirectlywithRNApolymeraseII. Methodology: Here we use co-localization, co-immunoprecipitation (co-IP) and in situ proximity ligation assay (PLA) to identifyBrahmaRelatedGene1(BRG1),theATP-drivenmotorofthehumanSWI-SNFchromatinremodelingcomplex,as anotherSAF-Ainteractionpartnerinmouseembryonicstem(mES)cells.WealsoemployRNAinterferencetoinvestigate functionalaspectsoftheSAF-A/BRG1interaction. PrincipalFindings:WefindthatendogenousSAF-AproteininteractswithendogenousBRG1proteininmEScells,andthat theinteractiondoesnotsolelydependonthepresenceofmRNA.Moreovertheinteractionremainsintactwhencellsare inducedtodifferentiate.FunctionalanalysesrevealthatdualdepletionofSAF-AandBRG1abolishesglobaltranscriptionby RNApolymeraseII,whilethenucleolarRNApolymeraseItranscriptionmachineryremainsunaffected. Conclusions:WedemonstratethatSAF-AinteractswithBRG1andthatbothcomponentsarerequiredforRNAPolymeraseII MediatedTranscription. Citation:Vizlin-HodzicD,RunnbergR,RymeJ,SimonssonS,SimonssonT(2011)SAF-AFormsaComplexwithBRG1andBothComponentsAreRequiredforRNA PolymeraseIIMediatedTranscription.PLoSONE6(12):e28049.doi:10.1371/journal.pone.0028049 Editor:MaryBryk,TexasAMUniversity,UnitedStatesofAmerica ReceivedSeptember9,2011;AcceptedOctober31,2011;PublishedDecember6,2011 Copyright:?2011Vizlin-Hodzicetal.Thisisanopen-accessarticledistributedunderthetermsoftheCreativeCommonsAttributionLicense,whichpermits unrest