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Role of histone H3 lysine 27 methylation in polycomb-group silencing
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A human PRC2 complex is purified and found to contain HMTase activity directed toward K27 of histone H3. Functional verification is obtained in Drosophila using a temperature-sensitive allele of E(Z); in this manner, enzymatic activity is pinpointed to this protein. Using chromatin immunoprecipitation, it is demonstrated that loss of E(Z) function abolishes K27 methylation of, and Pc binding to, the upstream regulatory region of Ubx.
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Cao R., Wang L., Wang H., Xia L., Erdjument-Bromage H., Tempst P., Jones R.S., Zhang Y. Role of histone H3 lysine 27 methylation in polycomb-group silencing. Science. 298:2002;1039-1043 A human PRC2 complex is purified and found to contain HMTase activity directed toward K27 of histone H3. Functional verification is obtained in Drosophila using a temperature-sensitive allele of E(Z); in this manner, enzymatic activity is pinpointed to this protein. Using chromatin immunoprecipitation, it is demonstrated that loss of E(Z) function abolishes K27 methylation of, and Pc binding to, the upstream regulatory region of Ubx.
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Histone methyltransferase activity of a Drosophila Polycomb group repressor complex
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A PRC2 complex containing ESC, E(Z), NURF-55 and SU(Z)12 is purified from Drosophila embryos, reconstituted in vitro, and found to display H3 K27-directed HMTase activity. Mutations in E(Z) are found to abolish HMTase activity. Using a transgene rescue assay in E(Z)-mutant flies, the authors demonstrate that E(Z) is required to maintain silencing of different HOX genes in imaginal discs.
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Muller J., Hart C.M., Francis N.J., Vargas M.L., Sengupta A., Wild B., Miller E.L., O'Connor M.B., Kingston R.E., Simon J.A. Histone methyltransferase activity of a Drosophila Polycomb group repressor complex. Cell. 111:2002;197-208 A PRC2 complex containing ESC, E(Z), NURF-55 and SU(Z)12 is purified from Drosophila embryos, reconstituted in vitro, and found to display H3 K27-directed HMTase activity. Mutations in E(Z) are found to abolish HMTase activity. Using a transgene rescue assay in E(Z)-mutant flies, the authors demonstrate that E(Z) is required to maintain silencing of different HOX genes in imaginal discs.
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Histone methyltransferase activity associated with a human multiprotein complex containing the Enhancer of Zeste protein
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A human PRC2 complex is isolated from 293 cells using epitope-tagged EED. The purified complex is found to methylate K9 and K27 of histone H3 in a manner dependent on an intact SET domain in EZH2. In pull-down assays using modified histone H3 peptides, it is demonstrated that methylated K27 provides a binding site for Pc.
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Kuzmichev A., Nishioka K., Erdjument-Bromage H., Tempst P., Reinberg D. Histone methyltransferase activity associated with a human multiprotein complex containing the Enhancer of Zeste protein. Genes Dev. 16:2002;2893-2905 A human PRC2 complex is isolated from 293 cells using epitope-tagged EED. The purified complex is found to methylate K9 and K27 of histone H3 in a manner dependent on an intact SET domain in EZH2. In pull-down assays using modified histone H3 peptides, it is demonstrated that methylated K27 provides a binding site for Pc.
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Using an enhancer-trap approach, a novel cis-regulatory element, the global control region, is identified in the mouse HoxD complex. The global control region is highly conserved over the course of evolution, and is found to regulate the expression of structurally and functionally unrelated genes over distances of several 100 kb. In line with previous studies on the β-globin locus control region, this study underlines the importance of regulatory master switches on top of gene-specific regulation.
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Spitz F., Gonzalez F., Duboule D. A global control region defines a chromosomal regulatory landscape containing the HoxD cluster. Cell. 113:2003;405-417 Using an enhancer-trap approach, a novel cis-regulatory element, the global control region, is identified in the mouse HoxD complex. The global control region is highly conserved over the course of evolution, and is found to regulate the expression of structurally and functionally unrelated genes over distances of several 100 kb. In line with previous studies on the β-globin locus control region, this study underlines the importance of regulatory master switches on top of gene-specific regulation.
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In silico prediction of 167 candidate PREs on the basis of the presence of clustered binding sites for Pho, GAF and Zeste. A subset of the predicted PREs are subsequently verified using chromatin immunoprecipitation and transgenic analysis. Scanning of the predicted PREs reveals three novel sequence motifs, potentially representing binding sites for DNA binding proteins.
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Ringrose L., Rehmsmeier M., Dura J.M., Paro R. Genome-wide prediction of Polycomb/Trithorax response elements in Drosophila melanogaster. Dev Cell. 5:2003;759-771 In silico prediction of 167 candidate PREs on the basis of the presence of clustered binding sites for Pho, GAF and Zeste. A subset of the predicted PREs are subsequently verified using chromatin immunoprecipitation and transgenic analysis. Scanning of the predicted PREs reveals three novel sequence motifs, potentially representing binding sites for DNA binding proteins.
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The Drosophila polycomb group gene pleiohomeotic encodes a DNA binding protein with homology to the transcription factor YY1
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