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Promoter context-specific positive and negative transcriptional regulation of Glucocorticoid Receptor target genes by coregulator GLP.

ABSTRACT: A549 lung cancer cells were not infected (NI), transfected by plasmids containing shGLP (sequence targeting GLP gene) or shNS (non-specific sequence). These cells were treated with dex to identify subsets of genes regulated by dex (hormone) and GLP (EHMT1). NI and shNS data were submitted on the GSE35962.

ORGANISM(S): Homo sapiens

PROVIDER: GSE94646 | GEO | 2017/06/13

SECONDARY ACCESSION(S): PRJNA371728

REPOSITORIES: GEO

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Promoter context-specific positive and negative transcriptional regulation of Glucocorticoid Receptor target genes by coregulator G9a

Project description:A549 lung cancer cells were not infected (NI), transfected by plasmids containing shG9a (sequence targeting G9a gene) or shNS (non-specific sequence). These cells were treated with dex to identify subsets of genes regulated by dex (hormone) and G9a (EHMT2) Cells (NI/shNS/shG9a) were treated with 100nM dex for 24h and total RNA was extracted to identify differentially expressed genes

2012-11-02 | E-GEOD-35962 | biostudies-arrayexpress

Promoter context-specific positive and negative transcriptional regulation of Glucocorticoid Receptor target genes by coregulator G9a

2012-11-02 | GSE35962 | GEO

Post-natal supply of GLP/Ehmt1 reverses Histone H3K9 dimethylation in cortical neuron isolated from adult Ehmt1∆/+ mice brain

Project description:EHMT1 haploinsufficiency causes Kleefstra syndrome (KS), a complex disorder of developmental delay and intellectual disability. EHMT1 encodes a lysine methyltransferase GLP and regulates histone H3 lysine 9 dimethylation (H3K9me2). Ehmt1 heterozygous mutant (Ehmt1∆/+) mice show KS-like abnormal behavioral phenotypes. Here, we examined the reversibility of decreased H3K9me2 by postnatal supply of GLP in post-mitotic neuron of Ehmt1∆/+ mice brain. For this purpose, we generated the mice harboring inducible Ehmt1 cDNA with CamKII-creER (Ehmt1 iTG-neuron mice) in Ehmt1∆/+ background. These mice expressed exogenous Ehmt1 in post-mitotic neuron upon Tamoxifen treatment. Using these mice, we sorted out NeuN+ cells from cortical region and performed Histone H3 K9-dimethyation ChIP analysis. Here we showed that Ehmt1 haploinsufficiency induces global reduction of euchromatic H3K9me2 especially in the euchromatic region of brain and that postnatal supply of GLP can reverse the diminished H3K9me2 phenotype, even in a postmitotic situation.

2021-06-09 | GSE162327 | GEO

Identification of proteins stably associated with G9a/GLP (EHMT2/EHMT1) complex of histone methyltransferases

Project description:G9a (EHMT2) and the G9a-like protein GLP (EHMT1) form a stable G9a/GLP heterodimer in embryonic stem cells and function cooperatively to establish and maintain the abundant repressive H3K9me2 modification, in addition to modifying several non-histone proteins. The G9a-dependent H3K9me2 is implicated in lineage-specific gene silencing and covers large chromosomal domains. While the mechanism of H3K9me2maintenance by G9a/GLP is known, how new patterns of this modification are established is not well understood. With this in mind, we used FLAG affinity purification of G9a under two different stringency conditions (150 and 300 mM NaCl) coupled with mass spectrometry to identify proteins stably associated with G9a/GLP, which could serve as potential recruiters of the complex to unmodified chromatin.

2016-03-16 | PXD003466 | Pride

GLP is critical for oogenesis exhibiting a G9A-independent role in transcriptional repression

Project description:GLP (EHMT1) functions as an H3K9me1 and H3K9me2 methyltransferase through its reportedly obligatory dimerization with G9A (EHMT2). Here, we investigated the role of GLP in oocyte and embryo development in comparison to G9A using oocyte-specific conditional knockout mouse models (G9a cKO, Glp cKO, G9a-Glp cDKO). Loss of GLP in oogenesis severely impairs oocyte maturation, fertilization and embryo development, resulting in lethality before embryonic day E12.5. In contrast, loss of G9A has a milder effect with a proportion of embryos producing viable offspring. The Glp cKO also showed loss of G9A protein and, hence, was phenotypically very similar to the G9a-Glp cDKO. H3K9me2 was equally depleted in all cKO genotypes, whereas H3K9me1 was decreased only in Glp cKO and G9a-Glp cDKO oocytes. Furthermore, the transcriptome, DNA methylome and proteome were markedly more affected in G9a-Glp cDKO than G9a cKO oocytes, demonstrating that in the absence of GLP there are widespread epigenetic and gene expression changes in the oocyte independent of H3K9me2. Gene dysregulation with coupled changes in DNA methylation suggest localised loss of chromatin repression, resulting in upregulated protein expression. Together, our findings demonstrate that GLP can function independently of G9A in the oocyte and is required for oocyte developmental competence.

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Context-dependent requirement of H3K9 dimethyltransferase activity during reprogramming to pluripotency

Project description:The methylation of histone 3 at lysine 9 (H3K9) is widely regarded as a major roadblock for cellular reprogramming and interference with associated methyltransferases such as EHMT1 and EHMT2 (also known as GLP and G9A) facilitates the derivation of induced pluripotent stem cells (iPSCs). In addition, activation of demethylases by ascorbic acid (AA) has become a common approach to facilitate the extensive epigenetic remodeling required for iPSC formation. The functional interaction between the H3K9 methylation machinery and AA-stimulated enzymes, and consequences of interfering with these pathways, remain insufficiently explored. Here, we show that reduction of EHMT1/2 activity paradoxically counteracts iPSC formation in an optimized reprogramming system in the presence of AA. At the cellular and molecular level, EHMT1/2 activity is required for efficient downregulation of somatic genes and transition into an epithelial state. Genetic interference with the demethylase KDM3B partially rescues the adverse effect of EHMT1/2 inhibition on iPSC formation. Transient inhibition of EHMT1/2 during reprogramming yields iPSCs that fail to efficiently give rise to viable mice, suggesting persistent molecular defects in these cells. Together, our observations document a more nuanced function of H3K9 methyltransferases during iPSC formation and suggest that the balancing of AA-stimulated enzymes by EHMT1/2 is required for efficient and error-free reprogramming to pluripotency.

2020-09-28 | GSE130490 | GEO

Derepression of inflammation-related genes link to microglia activation and neural maturation defect in a mouse model of Kleefstra Syndrome

Project description:Kleefstra syndrome (KS, also known as 9q.34.3 deletion syndrome) is a rare genetic disorder characterized by a developmental delay, abnormal behaviors and autism-like features. This syndrome is caused by haplo-insufficiency of the euchromatin histone methyltransferase 1 gene (EHMT1/GLP/KDM1D). This gene product, GLP is a methyltransferase responsible for mono- and di-methylation of lysine 9 on histone H3 N-terminal tail, which modulates epigenetic information. Ehmt1 heterozygous mutant (Ehmt1∆/+) mice show KS-like abnormal behavioral phenotypes and utilized as KS model mice. Here, we isolated nuclei from adult Ehmt1∆/+ mice cortex and analysed gene expression precisely by single nucleus RNA-seq analysis. It showed that many genes were up or down regulated in different cell types of Ehmt1∆/+ mice brain. Among them, inflammation associating genes are up-regulated in Ehmt1∆/+ mice neuronal cells. And this up-regulation was reversed by postnatal supply of GLP in post-mitotic neuron of Ehmt1∆/+ mice brain.

2021-06-11 | GSE162934 | GEO

GLP-1 induced transcriptomic profile in INS-1 cells

Project description:Aim: The purpose of this RNAseq study is to gain insight into the effect that Glucagon Like Peptide-1(GLP-1) or BMS-21 (11 amino acid GLP-1 mimetic) has on the transcriptomic profile of an insulin secreting pancreatic beta cell line INS-1 at 1h and 6h. Methods: The mRNA-focussed libraries (constructed using TruSeq RNA Library Prep Kit v2) of GLP-1 or BMS-21 treated INS-1 cells at 1h or 6h were generated by sequencing, in quadruplicate, using an Illumina Hiseq2000. De-multiplexing and FASTQ sequence data was generated using the Illumina CASAVA1.8.2 pipeline. The sequence reads that passed quality filters were analyzed at the transcript level using the following. Alignment using TopHat (2.0.12) + Bowtie (2.2.3.0) to the illumina rat iGenome (build 75 - Rnor_5.0 (GCA_000001895.3)), reads summarised using Subread-feature counts (1.4.6) and differential gene analysis using DESeq2 (1.6.3) within Rstudio (0.98.945). Results: GLP-1 treatment of INS-1 cells induced many transcriptomic changes in gene expression at both 1h and 6h relative to control at 1h and 6h. At 1h following GLP-1 treatment there were 674 genes differentially expressed (470 up-regulated and 204 down-regulated, p-adj <0.05). Further, at 6h following exposure to GLP-1 treatment there were 3192 genes differently expressed (1709 up-regulated and 1,483 down-regulated, p-adj <0.05). Comparison of GLP-1 and BMS-21 treatments revealed highly similar transcriptomic profiles at both the 1h and 6h timepoints. Conclusion: GLP-1 has a varied effect on the transcriptomic profile of INS-1 cells.

2018-07-28 | GSE102003 | GEO

Polysome profiling of glp-4 mutant animals

Project description:To determine the translational status of mRNAs in glp-4 mutant animals, we analyzed total and polysomal RNA levels by tiling arrays A total of 8 samples was analyzed (quadruplicates of glp-4_total and glp-4_polysomal, respectively)

2015-01-08 | E-GEOD-62860 | biostudies-arrayexpress

glp-4 L4

Project description:glp-4 L4 vs mixed stage reference Keywords: other

2003-12-31 | GSE718 | GEO