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This study carefully analyzes the association of methylated histone H3 (on K4, K9 and K27 residues), Polycomb group proteins Ezh2 and Enx, and RNA polymerase II with the X chromosomes during the pre-implantation development of the mouse embryo. It concludes that the paternal X-chromosome initiates imprinted X inactivation at the four- to eight-cell stage, on the basis of Xist RNA-coating and the recruitment of Polycomb proteins. These repressive marks are found to be erased, and X-linked genes to be re-expressed in the inner cell mass at the blastocyst stage, prior to the onset of random X-chromosome inactivation.
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Okamoto I., Otte A.P., Allis C.D., Reinberg D., and Heard E. Epigenetic dynamics of imprinted X inactivation during early mouse development. Science 303 (2004) 644-649. This study carefully analyzes the association of methylated histone H3 (on K4, K9 and K27 residues), Polycomb group proteins Ezh2 and Enx, and RNA polymerase II with the X chromosomes during the pre-implantation development of the mouse embryo. It concludes that the paternal X-chromosome initiates imprinted X inactivation at the four- to eight-cell stage, on the basis of Xist RNA-coating and the recruitment of Polycomb proteins. These repressive marks are found to be erased, and X-linked genes to be re-expressed in the inner cell mass at the blastocyst stage, prior to the onset of random X-chromosome inactivation.
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The authors exploit a previously established system of induction that enables Xist to be turned on, off and on again during the course of differentiation of embryonic stem cells. This study suggests that memory mechanisms can add to or substitute for the self-perpetuation of heterochromatin in the maintenance of the inactive status.
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Kohlmaier A., Savarese F., Lachner M., Martens J., Jenuwein T., and Wutz A. A chromosomal memory triggered by Xist regulates histone methylation in X inactivation. PLoS Biol 2 (2004) E171. The authors exploit a previously established system of induction that enables Xist to be turned on, off and on again during the course of differentiation of embryonic stem cells. This study suggests that memory mechanisms can add to or substitute for the self-perpetuation of heterochromatin in the maintenance of the inactive status.
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••], this report shows a transient pairing of the two Xics at the onset of X inactivation in differentiated female embryonic stem cells. The pairing occurs between days 2 and 4 of differentiation. During the same time-window, other parts of the X chromosomes are shown not to pair. Pairing occurs around the time of Xist upregulation on the elect inactive X, but prior to recruitment of Ezh2 and H3K27me3 enrichment. It is shown that transient pairing is affected in Tsix homozygous but not heterozygous mutants, and in Xite heterozygous mutants. By a transgenesis approach, it is also shown that ectopic interactions can compete with Xic-Xic pairing. The take-home message is that counting and/or choice involve a physical interaction between the two Xics.
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••], this report shows a transient pairing of the two Xics at the onset of X inactivation in differentiated female embryonic stem cells. The pairing occurs between days 2 and 4 of differentiation. During the same time-window, other parts of the X chromosomes are shown not to pair. Pairing occurs around the time of Xist upregulation on the elect inactive X, but prior to recruitment of Ezh2 and H3K27me3 enrichment. It is shown that transient pairing is affected in Tsix homozygous but not heterozygous mutants, and in Xite heterozygous mutants. By a transgenesis approach, it is also shown that ectopic interactions can compete with Xic-Xic pairing. The take-home message is that counting and/or choice involve a physical interaction between the two Xics.
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••], this report describes the transient co-localization between the two Xics at the onset of X inactivation. The co-localization is maximal around day 1.5 and 2 of differentiation in two different female embryonic stem cell lines. It is affected by a heterozygous deletion 3′ of Xist, which abolishes the expression of Tsix and includes Xite. Restoring Tsix expression in this mutant embryonic stem cell line restores pairing of the two Xics despite the remaining heterozygosity for Xite. Multicopy Xic YAC (yeast artificial chromosome) transgenes can pair with the endogenous Xic locus in differentiated male embryonic stem cells, whereas single copy transgenes cannot. Therefore, pairing correlates with the capacity for counting and/or choosing of the transgene. Finally, this study shows that the Xic is preferentially localized to the nuclear periphery and that co-localization also occurs at the nuclear periphery.
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••], this report describes the transient co-localization between the two Xics at the onset of X inactivation. The co-localization is maximal around day 1.5 and 2 of differentiation in two different female embryonic stem cell lines. It is affected by a heterozygous deletion 3′ of Xist, which abolishes the expression of Tsix and includes Xite. Restoring Tsix expression in this mutant embryonic stem cell line restores pairing of the two Xics despite the remaining heterozygosity for Xite. Multicopy Xic YAC (yeast artificial chromosome) transgenes can pair with the endogenous Xic locus in differentiated male embryonic stem cells, whereas single copy transgenes cannot. Therefore, pairing correlates with the capacity for counting and/or choosing of the transgene. Finally, this study shows that the Xic is preferentially localized to the nuclear periphery and that co-localization also occurs at the nuclear periphery.
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In this study, cell culture models of imprinted and random X-inactivation in embryonic stem cells, trophoblast stem cells, and embryonic fibroblasts are analyzed by chromatin immunoprecipitation (ChIP) for recruitment of the pre-initiation transcription complex at both the Xist and the Tsix promoters. A clear correlation is found between Xist expression and complex recruitment at the Xist P1 promoter, suggesting that Xist upregulation operates, at least in part, at the level of transcriptional activation. This study further analyzes in embryonic stem cells the effect of Tsix transcription on chromatin conformation at the Xist-Tsix locus. Tsix is found to be responsible for H3K4me2 deposition within the Xist gene. The opposite effect is reported in the region of overlap between Tsix exon 4 and Xist, suggesting that the sense-antisense overlap around the Xist promoter region might elicit a closed chromatin conformation.
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dcX mice. Analysis of the CpG methylation and the DNaseI hypersensitive site at the Xist promoter region were performed in both embryonic and extra-embryonic lineages. It is shown that disruption of Tsix impairs establishment of a repressive structure at the Xist promoter.
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