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Histone turnover marks the boundaries of cis-regulatory domains

ABSTRACT: Cellular memory is maintained at the Drosophila homeotic gene clusters by cis-regulatory elements who mechanism of action is unknown. We have examined chromatin dynamics in the Drosophila genome by measuring histone turnover levels at high resolution. Surprisingly, homeotic gene clusters display characteristic histone turnover profiles with conspicuous peaks at boundaries of cis-regulatory domains superimposed over regions of very low turnover. Peaks of histone turnover precisely correspond to nuclease hypersensitive sites for epigenetic silencing proteins. Our results suggest that epigenetic regulation is facilitated by histone turnover, which maintains continuous accessibility ov cis-regulatory DNA. Keywords: Chromatin affinity-purification on microarray

ORGANISM(S): Drosophila melanogaster

PROVIDER: GSE6234 | GEO | 2007/03/01

SECONDARY ACCESSION(S): PRJNA97603

REPOSITORIES: GEO

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Drosophila melanogaster

Project description:Histone turnover marks the boundaries of cis-regulatory domains

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H3K27 modifications define segmental regulatory domains in the Drosophila bithorax complex.

Project description:The bithorax complex (BX-C) in the fruit fly, Drosophila melanogaster, is a cluster of homeotic genes that determines the identities the body segments. Expression of these genes is governed by cis-regulatory domains, one for each parasegment, which are arranged on the chromosome in the order of the parasegments they affect. Stable repression of these domains depends on the functions of the Polycomb Group, including its ability to methylate lysine 27 of histone H3. To learn whether PcG proteins generate parasegment-specific chromatin signatures, we have used transgenes to mark and isolate nuclei from single parasegments. These nuclei were profiled for histone modifications and chromatin proteins. The H3K27me3 profiles across the BX-C in successive parasegments show a striking “stairstep” pattern that reveals sharp boundaries of the BX-C regulatory domains. The borders of H3K27me3 modification domains are sharp, and align precisely with binding sites for the CCCTC-binding protein (CTCF). H3K27ac is broadly enriched across active domains, in a pattern complementary to K27me3. These findings provide a molecular definition of the homeotic domains, and implicate precisely localized H3K27 modification as a central determinant of segment identity. Nuclei from single parasegments of the Drosophila embryo were isolated and ChIP-seq performed for H3K27me3, CTCF, H3K27ac, H3K4me3, Ph, Pol2, Pc, and Suz12. For each sample, two biological replicates were performed.

2014-08-14 | E-GEOD-55257 | biostudies-arrayexpress

H3K27 modifications define segmental regulatory domains in the Drosophila bithorax complex.

2014-08-14 | GSE55257 | GEO

Tiling path analysis of Drosophila Schneider 2 cultured cells to map forum domains

Project description:Forum domains are stretches of chromosomal DNA that are excised from eukaryotic chromosomes during their spontaneous non-random fragmentation. Mostly forum domains are of 50-200 kb in length, although larger domains, up to 500 - 700 kb, are also observed. <br><br>We performed a genome-wide mapping of forum domains termini in Drosophila Schneider 2 cultured cells using Drosophila melanogaster genomic tiling microarray consisting of overlapping 3 kb fragments. We found that forum domains contain clusters of several or many genes inside. The largest forum domains correspond to the coordinately expressed main clusters of homeotic genes inside BX-C and ANTP-C, cluster of histone genes and clusters of piRNAs. PRE/TRE and binding sites of transcription factors are often reside inside domains, and do not overlap with forum domains termini. The terminal regions of domains often correspond to chromosomal sites containing different classes of mobile elements, binding sites of SuUR protein and regions of intercalary heterochromatin. Our results indicate that forum domains correspond to big multi-gene chromosomal units some of which could be co-coordinately activated or repressed.

2010-11-01 | E-MEXP-2468 | biostudies-arrayexpress

Deciphering the cis-regulatory architecture of mammalian photoreceptors

Project description:In order to define the genomic targets of Crx, we carried out Crx chromatin-immunoprecipitation followed by massively parallel sequencing (ChIP-seq) of eight-week-old mouse retinas using the Solexa platform. Sequence reads were mapped to the genome and 'peaks' were identified. These data were subjected to extensive bioinformatic analysis. In addition, selected peaks were experimentally tested for cis-regulatory activity by electroporation as promoter-reporter fusions into living mouse retinas. Over 5,000 Crx-bound regions (CBRs) were identified throughout the mouse genome. Many of these clusters of CBRs occur specifically around photoreceptor genes. In fact, Crx directly regulates the majority of known photoreceptor transcription factors as well as most known photoreceptor disease genes. Cis-regulatory analysis revealed that individual CBRs contribute in a combinatorial fashion to the overall activity of the gene they control. In addition, multiple Crx-binding sites within CBRs cooperatively interact via precise spacing rules to generate optimal levels of transcription. Crx ChIP-Seq analysis of C57BL/6 and Nrl-/- retinas. IgG ChIP-Seq was performed as a control. 1-2 replicates each.

2010-06-24 | E-GEOD-20012 | biostudies-arrayexpress

Spatially varying cis-regulatory divergence in Drosophila embryos elucidates cis-regulatory logic

Project description:Spatially varying cis-regulatory divergence in Drosophila embryos elucidates cis-regulatory logic

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Replication-independent nucleosome turnover confers the functional basis of cis-regulatory elements in adult heart

Project description:Through integrated H2BGFP ChIP-seq data over multiple conditions and time points, as well as other related ChIP-seq and RNA-seq data in adult mouse heart. We observed genomewide and locus-specific H2B turnover in non-proliferating adult cardiomyocytes (Cms), and a majority of genomic regions with rapid turnover rates distributes on cis-regulatory elements, as well as these regions are associated with heart function, regulatory factors binding and their activities. Moreover, H2B turnover rate is always positively correlated with gene transcription level on enhancers, promoters and exons. The rapid turnover rates at cis-regulatory elements can be partially explained by the binding and co-binding of RNAP II and various transcription factors (TFs). Our further analysis revealed that not only rapid H2B turnover at cis-regulatory elements responses to heart disease, but also slow turnover at H3K27me3 sites functionally responses to heart disease with a different manner. More generally, we indicate that the binding and co-binding of DNA-binding proteins, such as TFs and epigenetic enzymes, all of them coupled with H2B turnover to modulate gene transcription and chromatin state.

2020-04-17 | GSE97876 | GEO

Genome-wide incorporation dynamics reveal distinct categories of turnover for the histone variant H3.3

Project description:We developed a system to study the DNA replication-independent turnover nucleosomes containing the histone variant H3.3 in mammalian cells. By measuring the genome-wide incorporation of H3.3 at different time points following epitope-tagged H3.3 expression, we find three categories of H3.3-nucleosome turnover in vivo: rapid turnover, intermediate turnover and, specifically at telomeres, slow turnover. Our data indicate that H3.3-containing nucleosomes at enhancers and promoters undergo a rapid turnover that is associated with active histone modification marks including H3K4me1, H3K4me3, H3K9ac, H3K27ac and the histone variant H2A.Z. The rate of turnover is negatively correlated with H3K27me3 at regulatory regions and with H3K36me3 at gene bodies. Examination of incorporation dynamics of histone variant H3.3

2013-10-23 | E-GEOD-51505 | biostudies-arrayexpress

Drosophila melanogaster

Project description:Histone turnover mediates epigenetic processes in Drosophila S2 cells

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Reconstructing and reprogramming the tumor propagating potential of glioblastoma stem-like cells: ChIP-seq

Project description:Developmental fate decisions are dictated by master transcription factors (TFs) that interact with cis-regulatory elements to direct transcriptional programs. Certain malignant tumors may also depend on a cellular hierarchy reminiscent of normal development but superimposed on underlying genetic aberrations. In glioblastoma (GBM), a subset of stem-like tumor- propagating cells (TPCs) appears to drive tumor progression and underlie therapeutic resistance, yet remain poorly understood. Here, we identify a core set of neurodevelopmental TFs (POU3F2, SOX2, SALL2, OLIG2) essential for GBM propagation. These TFs coordinately bind and activate TPC-specific regulatory elements, and are sufficient to fully reprogram differentiated GBM cells to ‘induced’ TPCs that recapitulate the epigenetic landscape and phenotype of native TPCs. We reconstruct a TF network model that highlights critical interactions and identifies novel therapeutic targets for eliminating TPCs. Our study establishes the epigenetic basis of a developmental hierarchy in a devastating malignancy, provides detailed insight into the underlying gene regulatory programs, and suggests attendant therapeutic strategies. Histone modification profiling for H3K27ac and transcription factors in glioblastoma cell line reprogramming

2014-04-10 | E-GEOD-54047 | biostudies-arrayexpress

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