CAF-1 Is Essential for Heterochromatin Organization in Pluripotent Embryonic Cells 英文参考文献.docVIP

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CAF-1 Is Essential for Heterochromatin Organization in Pluripotent Embryonic Cells 英文参考文献.doc

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CAF-1 Is Essential for Heterochromatin Organization in Pluripotent Embryonic Cells 英文参考文献

CAF-1IsEssentialforHeterochromatin OrganizationinPluripotentEmbryonicCells Martin Houlard1,2¤,Soizik Berlivet1[,Aline V.Probst2[,Jean-Pierre Quivy2,Patrick He′ry1,Genevie`ve Almouzni2, Matthieu Ge′rard1* 1 Epigenetic Regulation and Cancer Group, Service de Biologie Mole′culaire Syste′mique, De′partement de Biologie Joliot-Curie, Direction des Sciences du Vivant, Commissariata`l’EnergieAtomique,CEASaclay,Gif-sur-Yvette,France,2SectiondeRecherche,InstitutCurie,UMR218duCentreNationaldelaRechercheScientifique,Paris, France During mammalian development, chromatin dynamics and epigenetic marking are important for genome reprogramming. Recent data suggest an important role for the chromatin assembly machinery in this process. To analyzetheroleofchromatinassemblyfactor1(CAF-1)duringpre-implantationdevelopment,wegeneratedamouse linecarryingatargetedmutationinthegeneencodingitslargesubunit,p150CAF-1.Lossofp150CAF-1inhomozygous mutantsleadstodevelopmentalarrestatthe16-cellstage.Absenceofp150CAF-1intheseembryosresultsinsevere alterations in the nuclear organization of constitutive heterochromatin. We provide evidence that in wild-type embryos, heterochromatin domains are extensively reorganized between the two-cell and blastocyst stages. In p150CAF-1 mutant 16-cell stage embryos, the altered organization of heterochromatin displays similarities to the structureofheterochromatinintwo-tofour-cellstagewild-typeembryos,suggestingthatCAF-1isrequiredforthe maturationofheterochromatinduringpreimplantationdevelopment.Inembryonicstemcells,depletionofp150CAF-1 using RNA interference results in the mislocalization, loss of clustering, and decondensation of pericentric heterochromatin domains. Furthermore, loss of CAF-1 in these cells results in the alteration of epigenetic histone methylationmarksatthelevelofpericentricheterochromatin.Thesealterationsofheterochromatinarenotfoundin p150CAF-1-depletedmouseembryonicfibroblasts,whicharecellsthatarealreadylineagecommitted,suggestingtha