Exportin 4 Interacts with Sox9 through the HMG Box and Inhibits the DNA Binding of Sox9 英文参考文献.docVIP

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Exportin 4 Interacts with Sox9 through the HMG Box and Inhibits the DNA Binding of Sox9 英文参考文献.doc

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Exportin 4 Interacts with Sox9 through the HMG Box and Inhibits the DNA Binding of Sox9 英文参考文献

Exportin4InteractswithSox9throughtheHMGBoxand InhibitstheDNABindingofSox9 MegumiTsuchiya1,2,HidesatoOgawa1,2,3*,TaigaSuzuki4,NoriyukiSugiyama5,TokukoHaraguchi1,3, YasushiHiraoka1,3 * 1GraduateSchoolofFrontierBiosciences,OsakaUniversity,Suita,Japan,2DivisionofSexDifferentiation,NationalInstituteforBasicBiology,NationalInstitutesofNatural Sciences, Okazaki, Japan, 3Advanced ICT Research Institute Kobe, National Institute of Information and Communications Technology, Kobe, Japan, 4Neuroscience ResearchInstitute,UniversityofCaliforniaatSantaBarbara,SantaBarbara,California,UnitedStatesofAmerica,5KyotoPrefecturalUniversityofMedicine,Kawaramachi- dori,Kyoto,Japan Abstract Sox9 is a transcription factor that is required for tissue development in mammals. In general, such transcription factors require co-regulators for precise temporal and spatial control of the activation and inactivation of the numerous genes necessary for precise development during embryogenesis. Here we identify a new Sox9 co-regulator: Using affinity chromatographywithimmobilizedSox9protein,weidentifiedexportin4(Exp4)asaninteractingproteinofSox9inhuman culturedcells.InteractionbetweenendogenousExp4andSox9proteinswasconfirmedinthehumanosteosarcomaU2OS cells by immunoprecipitation experiments using anti-Sox9 antibody. siRNA depletion of Exp4 enhanced transcription of Sox9targetgenesinU2OScells,butdidnotaffectnuclearlocalizationofSox9.TheseresultssuggestthatExp4regulates Sox9activityinthenucleus.FurthermorewefoundthattheHMGboxofSox9wasresponsibleforbindingtoExp4,andthe HMGboxwasrequiredforsuppressionofSox9-mediatedtranscription.ThiscontrastswiththeknownSox9co-regulators whichbindtoitstranscriptionalactivationdomain.ChromatinimmunoprecipitationanalysesrevealedthatExp4prevents Sox9bindingtotheenhancersofitstargetgenes. TheseresultsdemonstratethatExp4actsasaSox9 co-regulatorthat directlyregulatesbindingofSox9toitstargetgenes. Citation: Tsuchiya M,Ogawa H, Suzuki T,SugiyamaN, Haraguchi T,et al.(2011