Elements of the Polycomb Repressor SU(Z)12 Needed for Histone H3-K27 Methylation, the Interface with E(Z), and <i>In Vivo</i> Function

Aswathy, N.; Vargas, Marcus L.; Wang, Liangjun; Andersen, Erica; Miller, E. L.; Simon, John

doi:10.1128/mcb.00307-13

Cited by 31 publications

(45 citation statements)

References 78 publications

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“…Recombinant Spt6 did not inhibit in vitro methyltransferase activity of preformed PRC2 complex (data not shown), suggesting that Spt6 may prevent PRC2 assembly rather than promoting its disassembly. In Drosophila melanogaster , deletion of the Suz12 region involved in Ezh2 and Spt6 interactions results in reduced PRC2 binding to chromatin targets and strong loss of function in genetic rescue tests (Rai et al, 2013). In summary, our findings indicate that, beside behaving as a histone chaperone and transcriptional elongation factor (Kwak and Lis, 2013), Spt6 has the potential of partitioning the genome in transcribed and repressed regions by counteracting formation and recruitment of PRC2 and H3K27me3 at defined regulatory regions.…”

Section: Discussionmentioning

confidence: 99%

The Elongation Factor Spt6 Maintains ESC Pluripotency by Controlling Super-Enhancers and Counteracting Polycomb Proteins

Wang

Juan

et al. 2017

Molecular Cell

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SUMMARY Spt6 coordinates nucleosome dis- and re-assembly, transcriptional elongation and mRNA processing. Here, we report that depleting Spt6 in ESCs reduced expression of pluripotency factors, increased expression of cell lineage-affiliated developmental regulators, and induced cell morphological and biochemical changes indicative of ESC differentiation. Selective down-regulation of pluripotency factors upon Spt6 depletion may be mechanistically explained by its enrichment at ESC super-enhancers where Spt6 controls histone H3K27 acetylation and methylation, and super-enhancer RNA transcription. In ESCs, Spt6 interacted with the PRC2 core subunit Suz12 and prevented H3K27me3 accumulation at ESC super-enhancers and associated promoters. Biochemical as well as functional experiments revealed that Spt6 could compete for binding of the PRC2 methyltransferase Ezh2 to Suz12 and reduce PRC2 chromatin engagement. Thus, in addition to serving as a histone chaperone and transcription elongation factor, Spt6 counteracts repression by opposing H3K27me3 deposition at critical genomic regulatory regions.

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Section: Discussionmentioning

confidence: 99%

The Elongation Factor Spt6 Maintains ESC Pluripotency by Controlling Super-Enhancers and Counteracting Polycomb Proteins

Wang

Juan

et al. 2017

Molecular Cell

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“…HMTase assays on mutant PRC2 complexes (Fig. 2C) were performed as described (RAI et al 2013) and were repeated at least twice using independent preparations of mutant PRC2. Histone content was visualized by Amido Black staining.…”

Section: Methodsmentioning

confidence: 99%

“…S2 cell transfections were performed as described (RAI et al 2013), except S2 cells were transfected twice, on days 1 and 4 of culture, prior to harvesting for assays on day 7. ChIP assays were performed as described (WANG et al 2010) using 5 µl anti-H3K27me1 (Active Motif), 5 µl anti-H3K27Ac…”

Section: S2 Cell Transfections Chromatin Immunoprecipitations (Chipsmentioning

confidence: 99%

A Role for Monomethylation of Histone H3-K27 in Gene Activity inDrosophila

et al. 2018

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Polycomb repressive complex 2 (PRC2) is a conserved chromatin-modifying enzyme that methylates histone H3 on lysine-27 (K27). PRC2 can add one, two, or three methyl groups and the fully methylated product, H3-K27me3, is a hallmark of Polycomb-silenced chromatin.Less is known about functions of K27me1 and K27me2 and the dynamics of flux through these states. These modifications could serve mainly as intermediates to produce K27me3 or they could each convey distinct epigenetic information. To investigate this, we engineered a variant of Drosophila melanogaster PRC2 which is converted into a mono-methyltransferase.

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“…Samples were incubated for 60 minutes at 30°C and stopped by addition of SDS loading buffer prior to loading on a 4-12% NuPage Bis-Tris gel (Invitrogen). Western blotting, amido black staining (Sigma) and autoradiography were performed as described 46 . Quantification of amino black signal and autoradiography films was performed using ImageJ software (NIH).…”

Section: Methodsmentioning

confidence: 99%

A strand-specific switch in noncoding transcription switches the function of a Polycomb/Trithorax response element

et al. 2014

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Polycomb/Trithorax response elements (PRE/TREs) can switch their function reversibly between silencing and activation, by mechanisms that are poorly understood. Here we show that a switch in forward and reverse noncoding transcription from the Drosophila vestigial (vg) PRE/TRE switches the status of the element between silencing (induced by the forward strand) and activation (induced by the reverse strand). In vitro, both ncRNAs inhibit PRC2 histone methyltransferase activity, but in vivo only the reverse strand binds PRC2. Over-expression of the reverse strand evicts PRC2 from chromatin and inhibits its enzymatic activity. We propose that interactions of RNAs with PRC2 are differentially regulated in vivo, allowing regulated inhibition of local PRC2 activity. Genome-wide analysis shows that strand switching of ncRNAs occurs at several hundred PcG binding sites in fly and vertebrate genomes. This work identifies a novel and potentially widespread class of PRE/TREs that switch function by switching the direction of ncRNA transcription.

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Elements of the Polycomb Repressor SU(Z)12 Needed for Histone H3-K27 Methylation, the Interface with E(Z), and In Vivo Function

Cited by 31 publications

References 78 publications

The Elongation Factor Spt6 Maintains ESC Pluripotency by Controlling Super-Enhancers and Counteracting Polycomb Proteins

The Elongation Factor Spt6 Maintains ESC Pluripotency by Controlling Super-Enhancers and Counteracting Polycomb Proteins

A Role for Monomethylation of Histone H3-K27 in Gene Activity inDrosophila

A strand-specific switch in noncoding transcription switches the function of a Polycomb/Trithorax response element

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