DNA Methylation Supports Intrinsic Epigenetic Memory in Mammalian Cells 英文参考文献.docVIP

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DNA Methylation Supports Intrinsic Epigenetic Memory in Mammalian Cells 英文参考文献.doc

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DNA Methylation Supports Intrinsic Epigenetic Memory in Mammalian Cells 英文参考文献

DNAMethylationSupportsIntrinsic EpigeneticMemoryinMammalianCells Yong-Qing Feng1,Romain Desprat1,Haiqing Fu2,Emmanuel Olivier1,Chii Mei Lin2,Amanda Lobell1,Shilpa N.Gowda2, Mirit I.Aladjem2,Eric E.Bouhassira1* 1 Department of Medicine, Division of Hematology, and Department of Cell Biology, Albert Einstein College Of Medicine, Bronx, New York, New York, United States of America,2LaboratoryofMolecularPharmacology,NationalCancerInstitute,Bethesda,Maryland,UnitedStatesofAmerica WehaveinvestigatedtheroleofDNAmethylationintheinitiationandmaintenanceofsilencedchromatininsomatic mammalian cells. We found that a mutated transgene, in which all the CpG dinucleotides have been eliminated, underwenttranscriptionalsilencingtothesameextentastheunmodifiedtransgene.Theseobservationsdemonstrate that DNA methylation is not required for silencing. The silenced CpG-free transgene exhibited all the features of heterochromatin, including silencing of transcriptional activity, delayed DNA replication, lack of histone H3 and H4 acetylation,lackofH3-K4methylation,andenrichmentintri-methyl-H3-K9.Incontrast,whenwetestedfortransgene reactivationusingaCrerecombinase-mediatedinversionassay,weobservedamarkeddifferencebetweenaCpG-free and an unmodified transgene: the CpG-free transgene resumed transcription and did not exhibit markers of heterochromatinwhereastheunmodifiedtransgeneremainedsilenced.ThesedataindicatethatmethylationofCpG residuesconferredepigeneticmemoryinthissystem.Theseresultsalsosuggestthatreplicationdelay,lackofhistone H3andH4acetylation,H3-K4methylation,andenrichmentintri-methyl-H3-K9arenotsufficienttoconferepigenetic memory. We propose that DNA methylation within transgenes serves as an intrinsic epigenetic memory to permanentlysilencetransgenesandpreventtheirreactivation. Citation:FengYQ,DespratR,FuH,OlivierE,LinCM,etal.(2006)DNAmethylationsupportsintrinsicepigeneticmemoryinmammaliancells.PLoSGenet2(4):e65.DOI:10. 1371/journal.pgen.0020065 associated with active chromati