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The exosome degrades chromatin-associated RNAs genome-wide and maintains chromatin homeostasis

ABSTRACT: This SuperSeries is composed of the SubSeries listed below.

ORGANISM(S): Drosophila melanogaster

PROVIDER: GSE222262 | GEO | 2025/01/01

REPOSITORIES: GEO

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The exosome degrades chromatin-associated RNAs genome-wide and maintains chromatin homeostasis (RNA-Seq)

Project description:Eukaryotic genomes produce a large variety of RNAs some of which are present in the chromatin and may affect important processes such as chromatin packaging and gene expression. Little is known about the role of the RNA exosome in the regulation of such chromatin-associated RNAs (caRNAs). The project aims at 1) identifying caRNAs produced from unique and repetitive sequences in the genome of Drosophila melanogaster S2 cells, and 2) characterize the role of the RNA exosome in controlling caRNA levels. To answer these questions, we have used RNA interference to knock down the exosome catalytic subunits RRP6 and DIS3 (individually or simultaneously) and we have analyzed the effects of the depletions on the chromatin-associated transcriptome by sequencing RNAs purified from a chromatin fraction isolated from S2 cells.

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The exosome degrades chromatin-associated RNAs genome-wide and maintains chromatin homeostasis (ATAC-Seq)

Project description:Eukaryotic genomes produce a large variety of RNAs some of which are present in the chromatin and may affect important processes such as chromatin packaging and gene expression. The RNA exosome controls the levels of chromatin-associated RNAs (caRNAs), but little is known about its role in the regulation of chromatin packaging. The project aims at characterizing the role of the RNA exosome in the maintenance of chromatin packaging in Drosophila melanogaster S2 cells. We have used RNA interference to knock down the exosome catalytic subunits RRP6 and DIS3 (individually or simultaneously) and we have analyzed the effects of the depletions on chromatin accessibility by ATAC-seq.

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The exosome degrades chromatin-associated RNAs genome-wide and maintains chromatin homeostasis

Project description:The exosome degrades chromatin-associated RNAs genome-wide and maintains chromatin homeostasis

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The exosome degrades chromatin-associated RNAs genome-wide and maintains chromatin homeostasis (ATAC-Seq)

Project description:The exosome degrades chromatin-associated RNAs genome-wide and maintains chromatin homeostasis (ATAC-Seq)

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The exosome degrades chromatin-associated RNAs genome-wide and maintains chromatin homeostasis (RNA-Seq)

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KDM1A maintains genome-wide homeostasis of transcriptional enhancers.

Project description:Transcriptional enhancers enable exquisite spatiotemporal control of gene expression in metazoans. Enrichment of monomethylation of histone H3 lysine 4 (H3K4me1) is a major chromatin signature of transcriptional enhancers. Lysine (K)-specific demethylase 1A (KDM1A, also known as LSD1), an H3K4me2/me1 demethylase, inactivates stem-cell enhancers during the differentiation of mouse embryonic stem cells (mESCs). However, its role in undifferentiated mESCs remains obscure. Here, we show that KDM1A actively maintains the optimal enhancer status in both undifferentiated and lineage-committed cells. KDM1A occupies a majority of enhancers in undifferentiated mESCs. KDM1A levels at enhancers exhibit clear positive correlations with its substrate H3K4me2, H3K27ac, and transcription at enhancers. In Kdm1a-deficient mESCs, a large fraction of these enhancers gains additional H3K4 methylation, which is accompanied by increases in H3K27 acetylation and increased expression of both enhancer RNAs (eRNAs) and target genes. In postmitotic neurons, loss of KDM1A leads to premature activation of neuronal activity-dependent enhancers and genes. Taken together, these results suggest that KDM1A is a versatile regulator of enhancers and acts as a rheostat to maintain optimal enhancer activity by counterbalancing H3K4 methylation at enhancers.

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Project description:This SuperSeries is composed of the SubSeries listed below.

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PHC1 maintains pluripotency by organizing genome-wide chromatin interactions of the Nanog locus.

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Genome-wide localization of exosome components to active promoters and chromatin insulators in Drosophila.

Project description:Chromatin insulators are functionally conserved DNA-protein complexes situated throughout the genome that organize independent transcriptional domains. Previous work implicated RNA as an important cofactor in chromatin insulator activity, although the precise mechanisms are not yet understood. Here we identify the exosome, the highly conserved major cellular 3' to 5' RNA degradation machinery, as a physical interactor of CP190-dependent chromatin insulator complexes in Drosophila. Genome-wide profiling of exosome by ChIP-seq in two different embryonic cell lines reveals extensive and specific overlap with the CP190, BEAF-32 and CTCF insulator proteins. Colocalization occurs mainly at promoters but also boundary elements such as Mcp, Fab-8, scs and scs', which overlaps with a promoter. Surprisingly, exosome associates primarily with promoters but not gene bodies of active genes, arguing against simple cotranscriptional recruitment to RNA substrates. Similar to insulator proteins, exosome is also significantly enriched at divergently transcribed promoters. Directed ChIP of exosome in cell lines depleted of insulator proteins shows that CTCF is required specifically for exosome association at Mcp and Fab-8 but not other sites, suggesting that alternate mechanisms must also contribute to exosome chromatin recruitment. Taken together, our results reveal a novel positive relationship between exosome and chromatin insulators throughout the genome.

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Functional interaction between the RNA exosome and the sirtuin deacetylase Hst3 maintains transcriptional homeostasis.

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