Article

CDYL Bridges REST and Histone Methyltransferases for Gene Repression and Suppression of Cellular Transformation

Department of Pathology, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, MA 02115, USA.
Molecular cell (Impact Factor: 14.46). 01/2009; 32(5):718-26. DOI: 10.1016/j.molcel.2008.10.025
Source: PubMed

ABSTRACT The neuronal gene repressor REST/NRSF recruits corepressors, including CoREST, to modify histones and repress transcription. REST also functions as a tumor suppressor, but the mechanism remains unclear. We identified chromodomain on Y-like (CDYL) as a REST corepressor that physically bridges REST and the histone methylase G9a to repress transcription. Importantly, RNAi knockdown of REST, CDYL, and G9a, but not CoREST, induced oncogenic transformation of immortalized primary human cells and derepression of the proto-oncogene TrkC. Significantly, transgenic expression of TrkC also induced transformation. This implicates CDYL-G9a, but not CoREST, in REST suppression of transformation, possibly by oncogene repression. CDYL knockdown also augments transformation in a cell culture model of cervical cancer, where loss of heterozygosity of the CDYL locus occurs. These findings demonstrate molecular strategies by which REST carries out distinct biological functions via different corepressors and provide critical insights into the role of histone-modifying complexes in regulating cellular transformation.

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Available from: Matthias Ottinger, Aug 29, 2015
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    • "None of the known epigenetic regulators identified as co-factors by mass spectrometry were specific to ESCs. However, we did identify almost all known REST co-factors including CoREST1 and Sin3a as well as the chromatin modifying enzymes, HDAC1 and 2, Kdm1a and G9a/Glp, and the G9a-associated adaptors CDYL and WIZ1, all of which have been shown biochemically to be present within REST complexes in terminally differentiated cell types (Andres et al., 1999; Grimes et al., 2000; Hakimi et al., 2002; Roopra et al., 2004; Mulligan et al., 2008), thus validating our approach. We noted that an additional CoREST family member, CoREST2, was also present in the pull-downs. "
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    ABSTRACT: The bivalent hypothesis posits that genes encoding developmentalregulators required for early lineage decisions are poised in stem/progenitor cells by the balance between a repressor histone modification (H3K27me3), mediated by the Polycomb Repressor Complex 2 (PRC2), and an activator modification (H3K4me3). Here, we test whether this mechanism applies equally to genes that are not required until terminal differentiation. We focus on the RE1 Silencing Transcription Factor (REST) because it is expressed highly in stem cells, and is an established global repressor of terminal neuronal genes. Elucidation of the REST complex, and comparison of chromatin marks and gene expression levels in control and REST-deficient stem cells, shows that REST target genes are poised by a mechanism independent of Polycomb, even at promoters which bear the H3K27me3 mark. Specifically, genes under REST control are actively repressed in stem cells by a balance of the H3K4me3 mark and a repressor complex that relies on histone deacetylase activity. Thus, chromatin distinctions between pro-neural and terminal neuronal genes are established at the embryonic stem cell stage by two parallel, but distinct, repressor pathways.
    eLife Sciences 09/2014; 3. DOI:10.7554/eLife.04235 · 8.52 Impact Factor
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    • "Namely, the Rest complex includes the histone deacetylase HDAC, which interacts with Rest through Sin3A and CoRest [19], [20]. The H3K4 demethylase Lsd1 and H3K9 methyl transferase G9a also interact through CoRest and Cdyl, respectively [21], [22], [23]. Such histone-modifying enzymes introduce repressive histone modifications to Rest binding sites. "
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    ABSTRACT: We detected and characterized the binding sites of the representative Rest complex components Rest, Sin3A, and Lsd1. We compared their binding patterns in mouse embryonic stem (ES) cells and epiblast stem (EpiS) cells. We found few Rest sites unique to the EpiS cells. The ES-unique site features were distinct from those of the common sites, namely, the signal intensities were weaker, and the characteristic gene function categories differed. Our analyses showed that the Rest binding sites do not always overlap with the Sin3A and Lsd1 binding sites. The Sin3A binding pattern differed remarkably between the ES and EpiS cells and was accompanied by significant changes in acetylated-histone patterns in the surrounding regions. A series of transcriptome analyses in the same cell types unexpectedly showed that the putative target gene transcript levels were not dramatically different despite dynamic changes in the Rest complex binding patterns and chromatin statuses, which suggests that Rest is not the sole determinant of repression at its targets. Nevertheless, we identified putative Rest targets with explicitly enhanced transcription upon Rest knock-down in 143 and 60 common and ES-unique Rest target genes, respectively. Among such sites, several genes are involved in ES cell proliferation. In addition, we also found that long, intergenic non-coding RNAs were apparent Rest targets and shared similar features with the protein-coding target genes. Interestingly, such non-coding target genes showed less conservation through evolution than protein-coding targets. As a result of differences in the components and targets of the Rest complex, its functional roles may differ in ES and EpiS cells.
    PLoS ONE 04/2014; 9(4):e95374. DOI:10.1371/journal.pone.0095374 · 3.23 Impact Factor
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    • "However, we cannot rule out the possibility that the MIER1α isoform is also shuttled out of the nucleus through interaction with a NES-containing protein. MIER1α has been shown to interact several molecules in addition to ERα [8] and HDAC1/2 [2]; these include the histone methyltransferase G9a [30], the chromodomain-containing protein CDYL [31] and the histone acetyltransferase CBP [3]. However, none of these has been reported to contain a NES. "
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    ABSTRACT: MIER1α is a transcriptional regulator that functions in gene repression through its ability to interact with various chromatin modifiers and transcription factors. We have also shown that MIER1α interacts with ERα and inhibits estrogen-stimulated growth. While MIER1α is localized in the nucleus of MCF7 cells, previous studies have shown that it does not contain a nuclear localization signal. In this report, we investigate the mechanism involved in transporting MIER1α into the nucleus. We explored the possibility that MIER1α is transported into the nucleus through a 'piggyback' mechanism. One obvious choice is via interaction with ERα, however we demonstrate that nuclear targeting of MIER1α does not require ERα. Knockdown of ERα reduced protein expression to 22% of control, but did not alter the percentage of cells with nuclear MIER1α (98% nuclear with scrambled shRNA vs. 95% with ERα shRNA). Further evidence was obtained using two stable transfectants derived from the ER-negative MDA231 cell line: MC2 (ERα+) and VC5 (ERα-). Confocal analysis showed no difference in MIER1α localization (86% nuclear in MC2 vs. 89% in VC5). These data demonstrate that ERα is not involved in nuclear localization of MIER1α. To identify the critical MIER1α sequence, we performed a deletion analysis and determined that the ELM2 domain was necessary and sufficient for nuclear localization. This domain binds HDAC1 & 2, therefore we investigated their role. Confocal analysis of an MIER1α containing an ELM2 point mutation previously shown to abolish HDAC binding revealed that this mutation results in almost complete loss of nuclear targeting: 10% nuclear vs. 97% with WT-MIER1α. Moreover, double knockdown of HDAC1 and 2 caused a reduction in percent nuclear from 86% to 44%. The results of this study demonstrate that nuclear targeting of MIER1α requires an intact ELM2 domain and is dependent on interaction with HDAC1/2.
    PLoS ONE 12/2013; 8(12):e84046. DOI:10.1371/journal.pone.0084046 · 3.23 Impact Factor
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