The Experts below are selected from a list of 81336 Experts worldwide ranked by ideXlab platform
Takashi Yokota - One of the best experts on this subject based on the ideXlab platform.
-
EED sox2 regulatory loop controls es cell self renewal through histone methylation and acetylation
The EMBO Journal, 2011Co-Authors: Hiroki Ura, Keita Kinoshita, Tadayuki Akagi, Hiroshi Koide, Kazuhiro Murakami, Shukuro Yamaguchi, Shinji Masui, Hitoshi Niwa, Takashi YokotaAbstract:Transcription factors and epigenetic modulators are involved in the maintenance of self-renewal in embryonic stem (ES) cells. Here, we demonstrate the existence of a regulatory loop in ES cells between Sox2, an indispensable transcription factor for self-renewal, and embryonic ectoderm development (EED), an epigenetic modulator regulating histone methylation. We found that Sox2 and EED positively regulate each other's expression. Interestingly, Sox2 overexpression suppressed the induction of differentiation-associated genes in EED-deficient ES cells without restoring histone methylation. This Sox2-mediated suppression was prevented by knockdown of the histone acetyltransferase (HAT), Tip60 or Elp3, and Sox2 stimulated expression of these HATs. Furthermore, forced expression of either HAT resulted in repression of differentiation-associated genes in EED-deficient cells. These results suggest that Sox2 overcame the phenotype of EED-deficient ES cells by promoting histone acetylation. We also found that knockout of EED and knockdown of these HATs synergistically enhanced the upregulation of differentiation-associated genes in ES cells. Taken together, our results suggest that the EED/Sox2 regulatory loop contributes to the maintenance of self-renewal in ES cells by controlling histone methylation and acetylation.
-
EED/Sox2 regulatory loop controls ES cell self‐renewal through histone methylation and acetylation
The EMBO journal, 2011Co-Authors: Hiroki Ura, Keita Kinoshita, Tadayuki Akagi, Hiroshi Koide, Kazuhiro Murakami, Shukuro Yamaguchi, Shinji Masui, Hitoshi Niwa, Takashi YokotaAbstract:Transcription factors and epigenetic modulators are involved in the maintenance of self-renewal in embryonic stem (ES) cells. Here, we demonstrate the existence of a regulatory loop in ES cells between Sox2, an indispensable transcription factor for self-renewal, and embryonic ectoderm development (EED), an epigenetic modulator regulating histone methylation. We found that Sox2 and EED positively regulate each other's expression. Interestingly, Sox2 overexpression suppressed the induction of differentiation-associated genes in EED-deficient ES cells without restoring histone methylation. This Sox2-mediated suppression was prevented by knockdown of the histone acetyltransferase (HAT), Tip60 or Elp3, and Sox2 stimulated expression of these HATs. Furthermore, forced expression of either HAT resulted in repression of differentiation-associated genes in EED-deficient cells. These results suggest that Sox2 overcame the phenotype of EED-deficient ES cells by promoting histone acetylation. We also found that knockout of EED and knockdown of these HATs synergistically enhanced the upregulation of differentiation-associated genes in ES cells. Taken together, our results suggest that the EED/Sox2 regulatory loop contributes to the maintenance of self-renewal in ES cells by controlling histone methylation and acetylation.
-
synergy of EED and tsix in the repression of xist gene and x chromosome inactivation
The EMBO Journal, 2008Co-Authors: Shinwa Shibata, Takashi Yokota, Anton WutzAbstract:X-chromosome inactivation (XCI) depends on the noncoding Xist gene. Xist transcription is negatively regulated by its antisense partner Tsix, whose disruption results in nonrandom XCI in females. However, males can maintain Xist in a repressed state without Tsix, indicating participation of additional factor(s) in the protection of the single male X from inactivation. Here, we provide evidence that the histone methyltransferase EED is also involved in the process. Male embryonic stem cells with EED-null and Tsix mutations (XΔY EED−/−) showed Xist hyperactivation upon differentiation, whereas cells with either mutation alone did not. Impaired X-linked gene expression was observed in the XΔY EED−/− ES cells at the onset of differentiation. The Xist promoter in the XΔY EED−/− cells showed elevated histone H3-dimethyl lysine 4 modifications and lowered CpG methylation, which are characteristics of open chromatin. Hence, we identified EED as an additional major player in the regulation of Xist expression. The synergy of Polycomb group proteins and antisense Tsix transcription in Xist gene regulation explains why males can repress Xist without Tsix.
-
Synergy of EED and Tsix in the repression of Xist gene and X‐chromosome inactivation
The EMBO journal, 2008Co-Authors: Shinwa Shibata, Takashi Yokota, Anton WutzAbstract:X-chromosome inactivation (XCI) depends on the noncoding Xist gene. Xist transcription is negatively regulated by its antisense partner Tsix, whose disruption results in nonrandom XCI in females. However, males can maintain Xist in a repressed state without Tsix, indicating participation of additional factor(s) in the protection of the single male X from inactivation. Here, we provide evidence that the histone methyltransferase EED is also involved in the process. Male embryonic stem cells with EED-null and Tsix mutations (XΔY EED−/−) showed Xist hyperactivation upon differentiation, whereas cells with either mutation alone did not. Impaired X-linked gene expression was observed in the XΔY EED−/− ES cells at the onset of differentiation. The Xist promoter in the XΔY EED−/− cells showed elevated histone H3-dimethyl lysine 4 modifications and lowered CpG methylation, which are characteristics of open chromatin. Hence, we identified EED as an additional major player in the regulation of Xist expression. The synergy of Polycomb group proteins and antisense Tsix transcription in Xist gene regulation explains why males can repress Xist without Tsix.
-
STAT3 and Oct-3/4 control histone modification through induction of EED in embryonic stem cells.
The Journal of biological chemistry, 2008Co-Authors: Hiroki Ura, Masayuki Usuda, Keita Kinoshita, Chuanhai Sun, Keitaro Mori, Tadayuki Akagi, Takahiko Matsuda, Hiroshi Koide, Takashi YokotaAbstract:Mouse embryonic stem (ES) cells can self-renew in the presence of leukemia inhibitory factor (LIF). Several essential transcription factors have been identified for the self-renewal of mouse ES cells, including STAT3, Oct-3/4, and Nanog. The molecular mechanism of ES cell self-renewal, however, is not fully understood. In the present study, we identified EED, a core component of Polycomb repressive complex 2, as a downstream molecule of STAT3 and Oct-3/4. Artificial activation of STAT3 resulted in increased expression of EED, whereas expression of a dominant negative mutant of STAT3 or suppression of Oct-3/4 expression led to down-regulation of EED. Reporter, chromatin immunoprecipitation, and electrophoretic mobility shift assays revealed that STAT3 and Oct-3/4 directly bind to the promoter region of EED, suggesting that EED is a common target molecule of STAT3 and Oct-3/4. We also found that suppression of STAT3, Oct-3/4, or EED causes induction of differentiation-associated genes as well as loss of Lys(27)-trimethylated histone H3 at the promoter regions of the differentiation-associated genes. Suppression of STAT3 and Oct-3/4 also resulted in the absence of EED at the promoter regions. These results suggest that STAT3 and Oct-3/4 maintain silencing of differentiation-associated genes through up-regulation of EED in self-renewing ES cells.
Anton Wutz - One of the best experts on this subject based on the ideXlab platform.
-
synergy of EED and tsix in the repression of xist gene and x chromosome inactivation
The EMBO Journal, 2008Co-Authors: Shinwa Shibata, Takashi Yokota, Anton WutzAbstract:X-chromosome inactivation (XCI) depends on the noncoding Xist gene. Xist transcription is negatively regulated by its antisense partner Tsix, whose disruption results in nonrandom XCI in females. However, males can maintain Xist in a repressed state without Tsix, indicating participation of additional factor(s) in the protection of the single male X from inactivation. Here, we provide evidence that the histone methyltransferase EED is also involved in the process. Male embryonic stem cells with EED-null and Tsix mutations (XΔY EED−/−) showed Xist hyperactivation upon differentiation, whereas cells with either mutation alone did not. Impaired X-linked gene expression was observed in the XΔY EED−/− ES cells at the onset of differentiation. The Xist promoter in the XΔY EED−/− cells showed elevated histone H3-dimethyl lysine 4 modifications and lowered CpG methylation, which are characteristics of open chromatin. Hence, we identified EED as an additional major player in the regulation of Xist expression. The synergy of Polycomb group proteins and antisense Tsix transcription in Xist gene regulation explains why males can repress Xist without Tsix.
-
Synergy of EED and Tsix in the repression of Xist gene and X‐chromosome inactivation
The EMBO journal, 2008Co-Authors: Shinwa Shibata, Takashi Yokota, Anton WutzAbstract:X-chromosome inactivation (XCI) depends on the noncoding Xist gene. Xist transcription is negatively regulated by its antisense partner Tsix, whose disruption results in nonrandom XCI in females. However, males can maintain Xist in a repressed state without Tsix, indicating participation of additional factor(s) in the protection of the single male X from inactivation. Here, we provide evidence that the histone methyltransferase EED is also involved in the process. Male embryonic stem cells with EED-null and Tsix mutations (XΔY EED−/−) showed Xist hyperactivation upon differentiation, whereas cells with either mutation alone did not. Impaired X-linked gene expression was observed in the XΔY EED−/− ES cells at the onset of differentiation. The Xist promoter in the XΔY EED−/− cells showed elevated histone H3-dimethyl lysine 4 modifications and lowered CpG methylation, which are characteristics of open chromatin. Hence, we identified EED as an additional major player in the regulation of Xist expression. The synergy of Polycomb group proteins and antisense Tsix transcription in Xist gene regulation explains why males can repress Xist without Tsix.
Terry Magnuson - One of the best experts on this subject based on the ideXlab platform.
-
Molecular and functional mapping of EED motifs required for PRC2-dependent histone methylation
Journal of molecular biology, 2007Co-Authors: Nathan D. Montgomery, Della Yee, Stephanie A. Montgomery, Terry MagnusonAbstract:Polycomb group proteins represent a conserved family of developmental regulators that mediate heritable transcriptional silencing by modifying chromatin states. One Polycomb group complex, the PRC2 complex, is composed of several proteins, including the histone H3 lysine 27 (H3K27) methyltransferase enhancer of zeste homolog 2 and the WD-repeat protein embryonic ectoderm development (EED). Histone H3K27 can be monomethylated (H3K27me1), dimethylated (H3K27me2), or trimethylated (H3K27me3). However, it remains unclear what regulates the number of methyl groups added to H3K27 in a particular nucleosome. In mammalian cells, EED is present as four distinct isoforms, which are believed to be produced by utilizing four distinct, in-frame translation start sites in a common EED mRNA. A mutation that disables all four EED isoforms produces defects in H3K27 methylation [Montgomery, N.D., Yee, D., Chen, A., Kalantry, S., Chamberlain, S.J., Otte, A.P. & Magnuson, T. (2005). The murine polycomb group protein EED is required for global histone H3 lysine-27 methylation. Curr. Biol., 15, 942-947]. To assess the roles of individual EED isoforms in H3K27 methylation, we first characterized three of the four EED isoform start sites and then demonstrated that individual isoforms are not necessary for H3K27me1, H3K27me2, or H3K27me3. Instead, we show that the core WD-40 motifs and the histone-binding region of EED alone are sufficient for the generation of all three marks, demonstrating that EED isoforms do not control the number of methyl groups added to H3K27.
-
The Polycomb group protein EED protects the inactive X-chromosome from differentiation-induced reactivation.
Nature cell biology, 2006Co-Authors: Sundeep Kalantry, Arie P Otte, Della Yee, Kyle C. Mills, Barbara Panning, Terry MagnusonAbstract:The Polycomb group (PcG) encodes an evolutionarily conserved set of chromatin-modifying proteins that are thought to maintain cellular transcriptional memory by stably silencing gene expression. In mouse embryos that are mutated for the PcG protein EED, X-chromosome inactivation (XCI) is not stably maintained in extra-embryonic tissues. EED is a component of a histone-methyltransferase complex that is thought to contribute to stable silencing in undifferentiated cells due to its enrichment on the inactive X-chromosome in cells of the early mouse embryo and in stem cells of the extra-embryonic trophectoderm lineage. Here, we demonstrate that the inactive X-chromosome in EED(-/-) trophoblast stem cells and in cells of the trophectoderm-derived extra-embryonic ectoderm in EED(-/-) embryos remain transcriptionally silent, despite lacking the PcG-mediated histone modifications that normally characterize the facultative heterochromatin of the inactive X-chromosome. Whereas undifferentiated EED(-/-) trophoblast stem cells maintained XCI, reactivation of the inactive X-chromosome occurred when these cells were differentiated. These results indicate that PcG complexes are not necessary to maintain transcriptional silencing of the inactive X-chromosome in undifferentiated stem cells. Instead, PcG proteins seem to propagate cellular memory by preventing transcriptional activation of facultative heterochromatin during differentiation.
-
Genome imprinting regulated by the mouse Polycomb group protein EED.
Nature genetics, 2003Co-Authors: Jesse Mager, Nathan D. Montgomery, Fernando Pardo-manuel De Villena, Terry MagnusonAbstract:Epigenetic regulation is essential for temporal, tissue-specific and parent-of-origin-dependent gene expression. It has recently been found that the mouse Polycomb group (PcG) gene EED (embryonic ectoderm development) acts to maintain repression of the imprinted X chromosome. Here, we investigated whether EED is also required for regulation of autosomal imprinted loci. Expression analyses showed that transcripts from the silent alleles of a subset of paternally repressed genes were present in EED(-/-) embryos. Parent-of-origin methylation was preserved in these embryos, but we observed changes in the methylation status of specific CpGs in differentially methylated regions (DMRs) at affected but not at unaffected loci. These data identify EED as a member of a new class of trans-acting factors that regulate parent-of-origin expression at imprinted loci.
-
The mouse PcG gene EED is required for Hox gene repression and extraembryonic development.
Mammalian genome : official journal of the International Mammalian Genome Society, 2002Co-Authors: Jianbo Wang, Jesse Mager, Elizabeth Schnedier, Terry MagnusonAbstract:The Polycomb group (PcG) of genes was first identified in Drosophila as maintenance factors for long-term transcriptional repression of homeotic genes. In mice, the PcG protein EED (Embryonic ectoderm development) is present in a distinct complex that interacts with histone deacetylase (HDAC) and the PcG member Ezh2 (Enhancer of zeste homolog 2), but not in the larger Polycomb repressive complex 1 (PRC1) formed by several other PcG proteins. EEDnull mutants manifest a distinct early gastrulation defect that occurs prior to homeotic gene expression. To determine whether EED is also required for regulating homeotic genes, a later acting EEDhypomorph mutation was analyzed. The anterior expression boundaries of several Hox genes were shifted rostrally by one segment, indicating that EED is required for stable repression of homeotic genes. Furthermore, although the EEDnull/hypomorph compound heterozygotes die during mid-gestation stage, they did not show a more severe derepression of Hox genes than the EEDhypomorph/hypomorph homozygotes. A detailed analysis of the mid-gestation lethality associated with the EEDnull/hypomorph compound heterozygotes revealed a novel function for EED in the development of secondary trophoblast giant cells during murine placenta formation. Tetraploid rescue experiments demonstrated that the defect is cell autonomous in the extraembryonic lineage. Mash2, a paternally imprinted gene important for trophoblast development, was ectopically expressed in the EED mutants. However, genetic crosses with a Mash2 null allele suggested that EED was not required to maintain Mash2 imprinting, but could be required in a lineage specific fashion to suppress Mash2 expression.
-
Cell and tissue requirements for the gene EED during mouse gastrulation and organogenesis.
Genesis (New York N.Y. : 2000), 2001Co-Authors: Elizabeth M. Morin-kensicki, Cynthia Faust, Christian Lamantia, Terry MagnusonAbstract:Mouse embryos homozygous for the allele EED(l7Rn5-3354SB) of the Polycomb Group gene embryonic ectoderm development (EED) display a gastrulation defect in which epiblast cells move through the streak and form extraembryonic mesoderm derivatives at the expense of development of the embryo proper. Here we demonstrate that homozygous mutant ES cells have the capacity to differentiate embryonic cell types both in vitro as embryoid bodies and in vivo as chimeric embryos. In chimeric embryos, EED mutant cells can respond to wild-type signals and participate in normal gastrulation movements. These results indicate a non-cell-autonomous function for EED. Evidence of mutant cell exclusion from the forebrain and segregation within somites, however, suggests that EED has cell-autonomous roles in aspects of organogenesis. A requirement for EED in the epiblast during embryonic development is supported by the fact that high-contribution chimeras could not be rescued by a wild-type extraembryonic environment.
Arie P Otte - One of the best experts on this subject based on the ideXlab platform.
-
The Polycomb group protein EED protects the inactive X-chromosome from differentiation-induced reactivation.
Nature cell biology, 2006Co-Authors: Sundeep Kalantry, Arie P Otte, Della Yee, Kyle C. Mills, Barbara Panning, Terry MagnusonAbstract:The Polycomb group (PcG) encodes an evolutionarily conserved set of chromatin-modifying proteins that are thought to maintain cellular transcriptional memory by stably silencing gene expression. In mouse embryos that are mutated for the PcG protein EED, X-chromosome inactivation (XCI) is not stably maintained in extra-embryonic tissues. EED is a component of a histone-methyltransferase complex that is thought to contribute to stable silencing in undifferentiated cells due to its enrichment on the inactive X-chromosome in cells of the early mouse embryo and in stem cells of the extra-embryonic trophectoderm lineage. Here, we demonstrate that the inactive X-chromosome in EED(-/-) trophoblast stem cells and in cells of the trophectoderm-derived extra-embryonic ectoderm in EED(-/-) embryos remain transcriptionally silent, despite lacking the PcG-mediated histone modifications that normally characterize the facultative heterochromatin of the inactive X-chromosome. Whereas undifferentiated EED(-/-) trophoblast stem cells maintained XCI, reactivation of the inactive X-chromosome occurred when these cells were differentiated. These results indicate that PcG complexes are not necessary to maintain transcriptional silencing of the inactive X-chromosome in undifferentiated stem cells. Instead, PcG proteins seem to propagate cellular memory by preventing transcriptional activation of facultative heterochromatin during differentiation.
-
establishment of histone h3 methylation on the inactive x chromosome requires transient recruitment of EED enx1 polycomb group complexes
Developmental Cell, 2003Co-Authors: Jose C R Silva, Arie P Otte, Winifred Mak, Ilona Zvetkova, Ruth Appanah, Tatyana B Nesterova, Zoe Webster, Antoine H F M Peters, Thomas Jenuwein, Neil BrockdorffAbstract:Previous studies have implicated the EED-Enx1 Polycomb group complex in the maintenance of imprinted X inactivation in the trophectoderm lineage in mouse. Here we show that recruitment of EED-Enx1 to the inactive X chromosome (Xi) also occurs in random X inactivation in the embryo proper. Localization of EED-Enx1 complexes to Xi occurs very early, at the onset of Xist expression, but then disappears as differentiation and development progress. This transient localization correlates with the presence of high levels of the complex in totipotent cells and during early differentiation stages. Functional analysis demonstrates that EED-Enx1 is required to establish methylation of histone H3 at lysine 9 and/or lysine 27 on Xi and that this, in turn, is required to stabilize the Xi chromatin structure.
-
the polycomb group protein EED interacts with yy1 and both proteins induce neural tissue in xenopus embryos
Molecular and Cellular Biology, 2001Co-Authors: David P E Satijn, Karien M Hamer, Jan Den L Blaauwen, Arie P OtteAbstract:Polycomb group (PcG) proteins form multimeric protein complexes which are involved in the heritable stable repression of genes. Previously, we identified two distinct human PcG protein complexes. The EED-EZH protein complex contains the EED and EZH2 PcG proteins, and the HPC-HPH PcG complex contains the HPC, HPH, BMI1, and RING1 PcG proteins. Here we show that YY1, a homolog of the Drosophila PcG protein pleiohomeotic (Pho), interacts specificially with the human PcG protein EED but not with proteins of the HPC-HPH PcG complex. Since YY1 and Pho are DNA-binding proteins, the interaction between YY1 and EED provides a direct link between the chromatin-associated EED-EZH PcG complex and the DNA of target genes. To study the functional significance of the interaction, we expressed the Xenopus homologs of EED and YY1 in Xenopus embryos. Both XEED and XYY1 induce an ectopic neural axis but do not induce mesodermal tissues. In contrast, members of the HPC-HPH PcG complex do not induce neural tissue. The exclusive, direct neuralizing activity of both the XEED and XYY1 proteins underlines the significance of the interaction between the two proteins. Our data also indicate a role for chromatin-associated proteins, such as PcG proteins, in Xenopus neural induction.
-
characterization of interactions between the mammalian polycomb group proteins enx1 ezh2 and EED suggests the existence of different mammalian polycomb group protein complexes
Molecular and Cellular Biology, 1998Co-Authors: Richard George Antonius Bernardus Sewalt, David P E Satijn, Karien M Hamer, Jan Den L Blaauwen, Johan Van Der Vlag, M J Gunster, Thijs Hendrix, Roel Van Driel, Arie P OtteAbstract:In Drosophila melanogaster, the Polycomb-group (PcG) and trithorax-group (trxG) genes have been identified as repressors and activators, respectively, of gene expression. Both groups of genes are required for the stable transmission of gene expression patterns to progeny cells throughout development. Several lines of evidence suggest a functional interaction between the PcG and trxG proteins. For example, genetic evidence indicates that the enhancer of zeste [E(z)] gene can be considered both a PcG and a trxG gene. To better understand the molecular interactions in which the E(z) protein is involved, we performed a two-hybrid screen with Enx1/EZH2, a mammalian homolog of E(z), as the target. We report the identification of the human EED protein, which interacts with Enx1/EZH2. EED is the human homolog of EED, a murine PcG gene which has extensive homology with the Drosophila PcG gene extra sex combs (esc). Enx1/EZH2 and EED coimmunoprecipitate, indicating that they also interact in vivo. However, Enx1/EZH2 and EED do not coimmunoprecipitate with other human PcG proteins, such as HPC2 and BMI1. Furthermore, unlike HPC2 and BMI1, which colocalize in nuclear domains of U-2 OS osteosarcoma cells, Enx1/EZH2 and EED do not colocalize with HPC2 or BMI1. Our findings indicate that Enx1/EZH2 and EED are members of a class of PcG proteins that is distinct from previously described human PcG proteins.
Shinwa Shibata - One of the best experts on this subject based on the ideXlab platform.
-
synergy of EED and tsix in the repression of xist gene and x chromosome inactivation
The EMBO Journal, 2008Co-Authors: Shinwa Shibata, Takashi Yokota, Anton WutzAbstract:X-chromosome inactivation (XCI) depends on the noncoding Xist gene. Xist transcription is negatively regulated by its antisense partner Tsix, whose disruption results in nonrandom XCI in females. However, males can maintain Xist in a repressed state without Tsix, indicating participation of additional factor(s) in the protection of the single male X from inactivation. Here, we provide evidence that the histone methyltransferase EED is also involved in the process. Male embryonic stem cells with EED-null and Tsix mutations (XΔY EED−/−) showed Xist hyperactivation upon differentiation, whereas cells with either mutation alone did not. Impaired X-linked gene expression was observed in the XΔY EED−/− ES cells at the onset of differentiation. The Xist promoter in the XΔY EED−/− cells showed elevated histone H3-dimethyl lysine 4 modifications and lowered CpG methylation, which are characteristics of open chromatin. Hence, we identified EED as an additional major player in the regulation of Xist expression. The synergy of Polycomb group proteins and antisense Tsix transcription in Xist gene regulation explains why males can repress Xist without Tsix.
-
Synergy of EED and Tsix in the repression of Xist gene and X‐chromosome inactivation
The EMBO journal, 2008Co-Authors: Shinwa Shibata, Takashi Yokota, Anton WutzAbstract:X-chromosome inactivation (XCI) depends on the noncoding Xist gene. Xist transcription is negatively regulated by its antisense partner Tsix, whose disruption results in nonrandom XCI in females. However, males can maintain Xist in a repressed state without Tsix, indicating participation of additional factor(s) in the protection of the single male X from inactivation. Here, we provide evidence that the histone methyltransferase EED is also involved in the process. Male embryonic stem cells with EED-null and Tsix mutations (XΔY EED−/−) showed Xist hyperactivation upon differentiation, whereas cells with either mutation alone did not. Impaired X-linked gene expression was observed in the XΔY EED−/− ES cells at the onset of differentiation. The Xist promoter in the XΔY EED−/− cells showed elevated histone H3-dimethyl lysine 4 modifications and lowered CpG methylation, which are characteristics of open chromatin. Hence, we identified EED as an additional major player in the regulation of Xist expression. The synergy of Polycomb group proteins and antisense Tsix transcription in Xist gene regulation explains why males can repress Xist without Tsix.