Active Chromatin Marks Are Retained on X Chromosomes Lacking Gene or Repeat Silencing Despite XISTXist Expression in Somatic Cell Hybrids 英文参考文献.docVIP

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Active Chromatin Marks Are Retained on X Chromosomes Lacking Gene or Repeat Silencing Despite XISTXist Expression in Somatic Cell Hybrids 英文参考文献.doc

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Active Chromatin Marks Are Retained on X Chromosomes Lacking Gene or Repeat Silencing Despite XISTXist Expression in Somatic Cell Hybrids 英文参考文献

ActiveChromatinMarksAreRetainedonX ChromosomesLackingGeneorRepeatSilencingDespite XIST/XistExpressioninSomaticCellHybrids NancyP.Thorogood,CarolynJ.Brown* DepartmentofMedicalGenetics,UniversityofBritishColumbia,Vancouver,Canada Abstract Background: X-chromosome inactivation occurs early in mammalian development and results in the inactive X chromosomeacquiringnumeroushallmarksofheterochromatin.WhileXISTisakeyplayerintheinactivationprocess,the methodofactionofthisncRNAisyettobedetermined. Methodology/PrincipalFindings:Toassesswhichfeaturesofheterochromatinmaybedirectlyrecruitedbytheexpression andlocalizationoftheXISTRNAwehaveanalyzedamouse/humansomaticcellhybridinwhichexpressionofhumanand mouseXIST/XisthasbeeninducedfromtheactiveXbydemethylation.Suchhybridshadpreviouslybeendemonstratedto disconnectXIST/XistexpressionfromgenesilencingandweconfirmmaintenanceofX-linkedgeneexpression,evencloseto theXistlocus,despitethelocalizedexpressionofmouseXist. Conclusions/Significance:LossoftheactivechromatinmarksH3acetylationandH3lysine4methylationwasnotobserved uponXIST/Xistexpression,norwasthereagainofDNAmethylation;thusthesemarksoffacultativeheterochromatinarenot solely dependent upon Xist expression. Cot-1 holes, regions of depleted RNA hybridization with a Cot-1 probe, were observeduponXistexpression;however,thesewereatreducedfrequencyandintensityinthesesomaticcells.Domainsof humanCot-1transcriptionwereobservedcorrespondingtothehumanchromosomesinthesomaticcellhybrids.TheCot-1 domain of theX was notreduced with the expressionof XIST, which fails tolocalize tothe human X chromosome in a mouse somatic cell background. The human inactive X in a mouse/human hybrid cell also shows delocalized XIST expressionandanongoingCot-1domain,despiteX-linkedgenesilencing.Theseresultsareconsistentwithrecentreports separating Cot-1 silencing from genic silencing, but also demonstrate repetitive element expression from an otherwise silentXchromosomeinthesehybridcells. Citation: Thorogood NP, Bro