Direct Identification of Insulator Components by Insertional Chromatin Immunoprecipitation 英文参考文献.docVIP

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Direct Identification of Insulator Components by Insertional Chromatin Immunoprecipitation 英文参考文献.doc

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Direct Identification of Insulator Components by Insertional Chromatin Immunoprecipitation 英文参考文献

DirectIdentificationofInsulatorComponentsby InsertionalChromatinImmunoprecipitation ToshitsuguFujita,HodakaFujii* CombinedProgramonMicrobiologyandImmunology,ResearchInstituteforMicrobialDiseases,OsakaUniversity,Osaka,Japan Abstract Comprehensive understanding of mechanisms of epigenetic regulation requires identification of molecules bound to genomicregionsofinterestinvivo.However,non-biasedmethodstoidentifymoleculesboundtospecificgenomiclociin vivoarelimited.Here,weappliedinsertionalchromatinimmunoprecipitation(iChIP)todirectidentificationofcomponents ofinsulatorcomplexes,whichfunctionasboundariesofchromatindomain.Wefoundthatthechickenb-globinHS4(cHS4) insulator complex contains an RNA helicase protein, p68/DDX5; an RNA species, steroid receptor RNA activator 1; and a nuclearmatrixprotein,Matrin-3,invivo.Bindingofp68andMatrin-3tothecHS4insulatorcoresequencewasmediatedby CCCTC-bindingfactor(CTCF).Thus,ourresultsshowedthatitisfeasibletodirectlyidentifyproteinsandRNAboundtoa specificgenomicregioninvivobyusingiChIP. Citation: Fujita T, Fujii H (2011) Direct Identification of Insulator Components by Insertional Chromatin Immunoprecipitation. PLoS ONE 6(10): e26109. doi:10.1371/journal.pone.0026109 Editor:RajaJothi,NationalInstitutesofHealth,UnitedStatesofAmerica ReceivedAugust12,2011;AcceptedSeptember19,2011;PublishedOctober17,2011 Copyright: ? 2011 Fujita, Fujii. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:ThisworkwassupportedbytheTokyoBiochemicalResearchFoundation(T.F.),http://www.tokyobrf.or.jp/english/;Grant-in-AidforYoungScientists(B) ((T.F.),Grant-in-AidforScientificResearchonInnovativeAreas((T.F.),((H.F.)fromtheMinistryofEducation,Culture,Sports, Science and Technology of Japan, http://www.mext.go.jp/english/a06.htm. The funders had no role in study