Evidence That Gene Activation and Silencing during Stem Cell Differentiation Requires a Transcriptionally Paused Intermediate State 英文参考文献.docVIP

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Evidence That Gene Activation and Silencing during Stem Cell Differentiation Requires a Transcriptionally Paused Intermediate State 英文参考文献.doc

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Evidence That Gene Activation and Silencing during Stem Cell Differentiation Requires a Transcriptionally Paused Intermediate State 英文参考文献

EvidenceThatGeneActivationandSilencingduring StemCellDifferentiationRequiresaTranscriptionally PausedIntermediateState JonathanL.Golob1,RoshanM.Kumar2,MatthewG.Guenther2,LilM.Pabon1,GabrielA.Pratt1,JeanneF. Loring3,LouiseC.Laurent3,4,RichardA.Young2,CharlesE.Murry1* 1DepartmentsofPathologyandBioengineering,CenterforCardiovascularBiology,InstituteforStemCellandRegenerativeMedicine,UniversityofWashington,Seattle, Washington,UnitedStatesofAmerica,2WhiteheadInstituteforBiomedicalResearch,Cambridge,Massachusetts,UnitedStatesofAmerica, 3CenterforRegenerative Medicine,TheScrippsResearchInstitute,LaJolla,California,UnitedStatesofAmerica,4DepartmentofReproductiveMedicine,UniversityofCaliforniaSanDiego,San Diego,California,UnitedStatesofAmerica Abstract A surprising portion of both mammalian and Drosophila genomes are transcriptionally paused, undergoing initiation withoutelongation.Wetestedthehypothesisthattranscriptionalpausingisanobligatetransitionstatebetweendefinitive activationandsilencingashumanembryonicstemcells(hESCs)changestatefrompluripotencytomesoderm.Chromatin immunoprecipitationfortrimethyllysine4onhistoneH3(ChIP-Chip)wasusedtoanalyzetranscriptionalinitiation,and39 transcriptarrayswereusedtodeterminetranscriptelongation.Pluripotentandmesodermalcellshadequivalentfractionsof thegenomeinactiveandpausedtranscriptionalstates(,48%each),with,4%definitivelysilenced(neitherinitiationnor elongation). Differentiation to mesoderm changed the transcriptional state of 12% of the genome, with roughly equal numbers of genes moving toward activation or silencing. Interestingly, almost all loci (98–99%) changing transcriptional statedosoeitherbyenteringorexitingthepausedstate.Amajorityofthesetransitionsinvolveeitherlossofinitiation,as genesspecifyingalternatelineagesarearchived,orgainofinitiation,inanticipationoffuturefull-lengthexpression.The additionofchromatindynamicspermittedmuchearlierpredictionsoffinalcellfatecomparedtosoleuseofconventional transcript arrays. The