CAF-1 is essential for heterochromatin organization in pluripotent embryonic cells.
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 analyze the role of chromatin assembly factor 1 (CAF-1) during pre-implantation development, we generated a mouse line carrying a targeted mutation in the gene encoding its large subunit, p150CAF-1. Loss of p150CAF-1 in homozygous mutants leads to developmental arrest at the 16-cell stage. Absence of p150CAF-1 in these embryos results in severe 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 structure of heterochromatin in two- to four-cell stage wild-type embryos, suggesting that CAF-1 is required for the maturation of heterochromatin during preimplantation development. In embryonic stem cells, depletion of p150CAF-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 methylation marks at the level of pericentric heterochromatin. These alterations of heterochromatin are not found in p150CAF-1-depleted mouse embryonic fibroblasts, which are cells that are already lineage committed, suggesting that CAF-1 is specifically required for heterochromatin organization in pluripotent embryonic cells. Our findings underline the role of the chromatin assembly machinery in controlling the spatial organization and epigenetic marking of the genome in early embryos and embryonic stem cells.
Houlard
Martin
M
Epigenetic Regulation and Cancer Group, Service de Biologie Moléculaire Systémique, Département de Biologie Joliot-Curie, Direction des Sciences du Vivant, Commissariat à l'Energie Atomique, CEA Saclay, Gif-sur-Yvette, France.
Berlivet
Soizik
S
Probst
Aline V
AV
Quivy
Jean-Pierre
JP
Héry
Patrick
P
Almouzni
Geneviève
G
Gérard
Matthieu
M
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D047108:Embryonic Development; D053595:Embryonic Stem Cells / Q000502:physiology*; D044127:Epigenesis, Genetic; D005091:Exons; D005095:Exoribonucleases; D005260:Female; D018390:Gene Targeting; D006570:Heterochromatin / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D039904:Pluripotent Stem Cells / Q000502:physiology*; D011506:Proteins / Q000235:genetics* / Q000502:physiology*; D012097:Repressor Proteins; D012260:Ribonucleases
D006570:Heterochromatin; D011506:Proteins; D012097:Repressor Proteins; C489778:Cnot7 protein, mouse; D005095:Exoribonucleases; D012260:Ribonucleases