Functional and Molecular Characterization of the Role of CTCF in Human Embryonic Stem Cell Biology 英文参考文献.docVIP

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Functional and Molecular Characterization of the Role of CTCF in Human Embryonic Stem Cell Biology 英文参考文献.doc

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Functional and Molecular Characterization of the Role of CTCF in Human Embryonic Stem Cell Biology 英文参考文献

FunctionalandMolecularCharacterizationoftheRoleof CTCFinHumanEmbryonicStemCellBiology SriKripaBalakrishnan1,MichaelWitcher1¤,TravisW.Berggren2,BeverlyM.Emerson1* 1RegulatoryBiologyLaboratory,SalkInstituteforBiologicalStudies,LaJolla,California,UnitedStatesofAmerica,2StemCellCore,SalkInstituteforBiologicalStudies,La Jolla,California,UnitedStatesofAmerica Abstract TheCCCTC-bindingfactorCTCFistheonlyknownvertebrateinsulatorproteinandhasbeenshowntoregulateimportant developmental processes such as imprinting, X-chromosome inactivation and genomic architecture. In this study, we examinedtheroleofCTCFinhumanembryonicstemcell(hESC)biology.WedemonstratethatCTCFassociateswithseveral importantpluripotencygenes,includingNANOG,SOX2,cMYCandLIN28andiscriticalforhESCproliferation.CTCFdepletion impactsexpressionofpluripotencygenesandaccelerateslossofpluripotencyuponBMP4induceddifferentiation,butdoes not result in spontaneous differentiation. We find that CTCF associates with the distal ends and internal sites of the co- regulated 160kb NANOG-DPPA3-GDF3 locus. Each of these sites can function as a CTCF-dependent enhancer-blocking insulatorinheterologousassays.InhESCs,CTCFexistsinmultisubunitproteincomplexesandcanbepoly(ADP)ribosylated. Known CTCF cofactors, such as Cohesin, differentially co-localize in the vicinity of specific CTCF binding sites within the NANOGlocus.Importantly,theassociationofsomecofactorsandproteinPARlationselectivelychangesupondifferentiation althoughCTCFbindingremainsconstant.UnderstandinghowuniquecofactorsmayimpartspecializedfunctionstoCTCFat specificgenomiclocationswillfurtherilluminateitsroleinstemcellbiology. Citation:BalakrishnanSK,WitcherM,BerggrenTW,EmersonBM(2012)FunctionalandMolecularCharacterizationoftheRoleofCTCFinHumanEmbryonicStem CellBiology.PLoSONE7(8):e42424.doi:10.1371/journal.pone.0042424 Editor:RajaJothi,NationalInstitutesofHealth,UnitedStatesofAmerica ReceivedApril10,2011;AcceptedJuly9,2012;PublishedAugust3,2012 Copyright:?2012Balakrishnan