Functional Consequences of Necdin Nucleocytoplasmic Localization 英文参考文献.docVIP

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Functional Consequences of Necdin Nucleocytoplasmic Localization 英文参考文献.doc

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Functional Consequences of Necdin Nucleocytoplasmic Localization 英文参考文献

FunctionalConsequencesofNecdinNucleocytoplasmic Localization AnatLavi-Itzkovitz1,MariannaTcherpakov1,ZehavaLevy1,ShalevItzkovitz1,FrancoiseMuscatelli2, MikeFainzilber1* 1DepartmentofBiologicalChemistry,WeizmannInstituteofScience,Rehovot,Israel,2InstitutdeNeurobiologiedelaMe′diterrane′e,INSERMU901,ParcScientifiquede LuminyBP13,Marseille,France Abstract Background:Necdin,aMAGEfamilyproteinexpressedprimarilyinthenervoussystem,hasbeenshowntointeractwith bothnuclearandcytoplasmicproteins,butthemechanismofitsnucleocytoplasmictransportareunknown. Methodology/PrincipalFindings:Wecarriedoutalarge-scaleinteractionscreenusingnecdinasabaitintheyeastRRS system, and found a wide range of potential interactors with different subcellular localizations, including over 60 new candidatesfordirectbindingtonecdin.Integrationoftheseinteractionsintoacomprehensivenetworkrevealedanumber of coherent interaction modules, including a cytoplasmic module connecting to necdin through huntingtin-associated protein1(Hap1),dynactinandhip-1proteininteractor(Hippi);anuclearP53andCreb-binding-protein(Crebbp)module, connectingthroughCrebbpandWWdomain-containingtranscriptionregulatorprotein1(Wwtr1);andanucleocytoplasmic transport module, connecting through transportins 1 and 2. We validated the necdin-transportin1 interaction and characterized a sequence motif in necdin that modulates karyopherin interaction. Surprisingly, a D234P necdin mutant showedenhancedbindingtobothtransportin1andimportinb1.Finally,exclusionofnecdinfromthenucleustriggered extensivecelldeath. Conclusions/Significance: These data suggest that necdin has multiple roles within protein complexes in different subcellular compartments, and indicate that it can utilize multiple karyopherin-dependent pathways to modulate its localization. Citation:Lavi-ItzkovitzA,TcherpakovM,LevyZ,ItzkovitzS,MuscatelliF,etal.(2012)FunctionalConsequencesofNecdinNucleocytoplasmicLocalization.PLoS ONE7(3):e33786.doi:10.1371/journal.pone.0033786 Editor:Christ