Modulation of the β-Catenin Signaling Pathway by the Dishevelled-Associated Protein Hipk1 英文参考文献.docVIP

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Modulation of the β-Catenin Signaling Pathway by the Dishevelled-Associated Protein Hipk1 英文参考文献.doc

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Modulation of the β-Catenin Signaling Pathway by the Dishevelled-Associated Protein Hipk1 英文参考文献

Modulationoftheb-CateninSignalingPathwaybythe Dishevelled-AssociatedProteinHipk1 SarahH.Louie1¤a,XiaoYongYang2,WilliamH.Conrad1,JeanotMuster1,StephaneAngers1¤b,RandallT. Moon1,BenjaminN.R.Cheyette2* 1HowardHughesMedicalInstitute,DepartmentofPharmacology,andInstituteforStemCellandRegenerativeMedicine,UniversityofWashingtonSchoolofMedicine, Seattle,Washington,UnitedStatesofAmerica,2DepartmentofPsychiatry,andGraduatePrograminDevelopmentalBiology,PrograminBiologicalSciences,Universityof CaliforniaSanFrancisco,SanFrancisco,California,UnitedStatesofAmerica Abstract Background: Wnts are evolutionarily conserved ligands that signal through b-catenin-dependent and b-catenin– independent pathways to regulate cell fate, proliferation, polarity, and movements during vertebrate development. Dishevelled (Dsh/Dvl) is a multi-domain scaffold protein required for virtually all known Wnt signaling activities, raising interestintheidentificationandfunctionsofDsh-associatedproteins. Methodology: We conducted a yeast-2-hybrid screen using an N-terminal fragment of Dsh, resulting in isolation of the XenopuslaevisorthologofHipk1.InteractionbetweentheDshandHipk1proteinswasconfirmedbyco-immunoprecipitation assaysandmassspectrometry,andfurtherexperimentssuggestthatHipk1alsocomplexeswiththetranscriptionfactorTcf3. SupportinganuclearfunctionduringX.laevisdevelopment,Myc-taggedHipk1localizesprimarilytothenucleusinanimal cap explants, and the endogenous transcript is strongly expressed during gastrula and neurula stages. Experimental manipulationsofHipk1levelsindicatethatHipk1canrepressWnt/b-catenintargetgeneactivation,asdemonstratedbyb- cateninreporterassaysinhumanembryonickidneycellsandbyindicatorsofdorsalspecificationinX.laevisembryosatthe lateblastulastage.Inaddition,asubsetofWnt-responsivegenessubsequentlyrequiresHipk1foractivationintheinvoluting mesodermduringgastrulation.Moreover,eitherover-expressionorknock-downofHipk1leadstoperturbedconvergent extensioncellmovementsinvolvedinbothgast