Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

CpG island-mediated global gene regulatory modes in mouse embryonic stem cells

ABSTRACT: In order to identify the regions occupied by transcription factors, we performed bio-ChIP followed by massive parallel sequencing (bioChIP-Seq) in mouse ES cells. A total of seven DNA transcription factors including Myc module (Myc, Max, E2F4, Tip60 and Dmap1) and Core module transcription factors (Nanog, Dax1; Kim et al., 2010) were identified. We show that the transcription factors can be classified into several classes according to their occupancies. Our data also show transcription factors in the same class co-occupy the same regulatory elements, share the common characteristics and act together. Identification of seven transcription factor binding sites in mES cells

ORGANISM(S): Mus musculus

SUBMITTER: Jonghwan Kim

PROVIDER: E-GEOD-48666 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

CpG island-mediated global gene regulatory modes in mouse embryonic stem cells.

Beck Samuel S Lee Bum-Kyu BK Rhee Catherine C Song Jawon J Woo Andrew J AJ Kim Jonghwan J

Nature communications 20141118

Both transcriptional and epigenetic regulations are fundamental for the control of eukaryotic gene expression. Here we perform a compendium analysis of >200 large sequencing data sets to elucidate the regulatory logic of global gene expression programs in mouse embryonic stem (ES) cells. We define four major classes of DNA-binding proteins (Core, PRC, MYC and CTCF) based on their target co-occupancy, and discover reciprocal regulation between the MYC and PRC classes for the activity of nearly al ...[more]

PMID: 25405324

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Project description:In mammalian cells transcription factors (TFs) preferentially bind sites contained in regions of high nucleosomal occupancy, as determined by nucleotide-dependent computational analysis. This observation suggests that nucleosomes may act as gatekeepers of TF binding sites. We hypothesized that in mammalian genomes the information controlling nucleosome assembly may partially coincide with the information that enables TFs to recognize cognate sites in cis-regulatory elements while ignoring the myriad of non-functional, randomly occurring consensus binding sites. This way, nucleosome-mediated masking would be coupled to TF binding site functionality. The hematopoietic master regulator Pu.1 maintained nucleosome depletion at macrophage-specific enhancers that were otherwise occupied by nucleosomes in other cell types and in reconstituted chromatin. We identified a minimal set of DNA sequence and shape features that predicted Pu.1 binding with 78% accuracy. The same features predicted nucleosome occupancy in cells where Pu.1 was not expressed with higher accuracy than specifically designed models. Control of nucleosome deposition by DNA sequence and shape features that also specify TF consensus site functionality may allow maintaining the gatekeeper function of nucleosomes during evolution of cis-regulatory elements. Chromatin immuno-precipitations of the transcription factor Pu.1 followed by multiparallel sequencing performed in murine bone marrow-derived macrophages. Experiments carried out in cells infected either with a retroviral vector containing a short hairpin targeting Pu.1 or with the empty vector as control. The shPU.1 hairpin (sequence available upon request) was selected among five designed using a publicly available software (http://katahdin.mssm.edu/siRNA) and was cloned in a modified version of TtRMPVIR inducible retroviral vector (Genbank HQ456318) in which the puromycin resistance gene was inserted. The empty vector, containing an sh-Renilla sequence, was used as control. Chromatin immuno-precipitations of the transcription factor PU.1 binding to in vitro reconstituted chromatin followed by multiparallel sequencing Micrococcal nuclease digestion of chromatin extracted from bone marrow derived macrophages followed by multiparallel sequencing . Experiments were carried out in untreated cells (4 replicates) and cells infected either with a retroviral vector containing a short hairpin targeting Pu.1 or with the empty vector as control (2 replicates). The shPU.1 hairpin (sequence available upon request) was selected among five designed using a publicly available software (http://katahdin.mssm.edu/siRNA) and was cloned in a modified version of TtRMPVIR inducible retroviral vector (Genbank HQ456318) in which the puromycin resistance gene was inserted. The empty vector, containing an sh-Renilla sequence, was used as control. Micrococcal nuclease digestion of in vitro reconstituted chromatin followed by multiparallel sequencing

Dataset Information

CpG island-mediated global gene regulatory modes in mouse embryonic stem cells

Publications

CpG island-mediated global gene regulatory modes in mouse embryonic stem cells.

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