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Cell-type specific and combinatorial usage of diverse transcription factors revealed by genome-wide binding studies in multiple human cells [new ChIP-Seq samples]

ABSTRACT: Cellular diversity in a multicellular organism like the human is achieved in part by distinct programs of gene expression at the level of transcription, which in turn are mediated by transcription factors (TFs). However, there are few systematic studies of the genomic binding of different types of TFs across a wide range of human cell types, especially in relation to gene expression. Using chromatin Immunoprecipitation followed by thigh-throughput sequencing (ChIP-seq) we identified an average of 45,000, 30,000 and 8,000 binding sites for CTCF, Pol2 and MYC respectively across eleven cell types.CTCF preferred to bind to intergenic regions while Pol2 and MYC tended to bind to core promoter regions. CTCF sites were highly conserved across diverse cell types, whereas MYC showed the greatest cell-type specificity. MYC co-localized with Pol2 at many of their binding sites and putative target genes. Cell-type specific binding sites, in particular for MYC and Pol II, were associated with cell-type specific functions. Patterns of binding in relation to gene features were generally invariant across different cell types. Pol II occupancy was higher over exons than adjacent introns, likely reflecting a link between transcriptional elongation and splicing. TF binding was positively correlated with the expression levels of their putative target genes, but combinatorial binding, in particular of MYC and Pol II was even more strongly associated with higher gene expression. Our ChIP-seq data sheds considerable light on how these transcription factors of different types function individually and in combination with one another to occupy target genomic loci and shape gene expression programs in a cell-type specific manner. For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf Keywords: Genome binding/occupancy profiling by high throughput sequencing

ORGANISM(S): Homo sapiens

PROVIDER: GSE32692 | GEO | 2011/10/18

REPOSITORIES: GEO

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Project description:Cellular diversity in a multicellular organism like the human is achieved in part by distinct programs of gene expression at the level of transcription, which in turn are mediated by transcription factors (TFs). However, there are few systematic studies of the genomic binding of different types of TFs across a wide range of human cell types, especially in relation to gene expression. Using chromatin Immunoprecipitation followed by thigh-throughput sequencing (ChIP-seq) we identified an average of 45,000, 30,000 and 8,000 binding sites for CTCF, Pol2 and MYC respectively across eleven cell types.CTCF preferred to bind to intergenic regions while Pol2 and MYC tended to bind to core promoter regions. CTCF sites were highly conserved across diverse cell types, whereas MYC showed the greatest cell-type specificity. MYC co-localized with Pol2 at many of their binding sites and putative target genes. Cell-type specific binding sites, in particular for MYC and Pol II, were associated with cell-type specific functions. Patterns of binding in relation to gene features were generally invariant across different cell types. Pol II occupancy was higher over exons than adjacent introns, likely reflecting a link between transcriptional elongation and splicing. TF binding was positively correlated with the expression levels of their putative target genes, but combinatorial binding, in particular of MYC and Pol II was even more strongly associated with higher gene expression. Our ChIP-seq data sheds considerable light on how these transcription factors of different types function individually and in combination with one another to occupy target genomic loci and shape gene expression programs in a cell-type specific manner. For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf Keywords: Genome binding/occupancy profiling by high throughput sequencing ChIP-seq were performed on eleven human cell lines: GM12878 (lymphoblastoid), K562 (leukemia), HepG2 (hepatocellular carcinoma), HelaS3 (cervical carcinoma), HUVEC (human umbilical vein endothelial cells), NHEK (keratinocytes), H1-ES (embryonic stem cells), MCF7 (breast adenocarcinoma), FB8470 (Normal child fibroblasts), FB0167P (Progeria fibroblast), and H54 (glioblastoma). For each cell line, two or three replicates were independently grown and split into three, one for each of the three experimental methods. Control ChIP experiments were performed on five of the cell lines with NHEK and H1-ES being excluded due to lack of material. Additional ChIP-Seq data mentioned in the 'overall design' but not included in this Series are contained in the GSE30226 SubSeries.

Project description:Cell-type specific transcriptional profiling is key to understanding cell fate specification and function. In order to achieve this it has been necessary, to date, to isolate specific cell types from complex tissues. We have developed 'TaDa', a technique that enables cell-specific profiling without cell isolation. TaDa permits genome-wide profiling of DNA- or chromatin-binding proteins without cell sorting, fixation or affinity purification. The method is simple, sensitive, highly reproducible and is in principle transferable to any model system. Here we show that TaDa can be used to identify transcribed genes in a cell-type specific manner. We profile the genome-wide binding of RNA polymerase II (Pol II) in adjacent, clonally related neural stem cells in intact Drosophila brains. Our data reveal the activity of non-canonical metabolic pathways in proliferating neuroepithelial cells, and highlight a possible role for the retinal determination gene regulatory network in patterning neural stem cell fates. We also identify temporal differences in the activity of signalling pathways that control neuroepithelial cell fate by profiling Pol II occupancy at two different stages of brain development. Using RNA Pol II TaDa to profile, in a cell-type specific manner, the transcriptional state of neuroepithelial cells at two stages of larval brain development. Closely related asymmetrically dividing neural stem cells (neuroblasts) were also profiled, in order to compare the transcriptomes of two different types of neural stem cells. 3 biological relicates were performed for 3rd instar neuroepithelial cells (with one dye-swap). 2 biological relicates were performed for 3rd instar neuroblasts (with dye-swap). 2 biological relicates were performed for 1st instar neuroepithelial cells (with dye-swap). As additional supporting evidence for the Pol II TaDa technique, 2 biological relicates were performed for 3rd instar salivary glands (with dye-swap) in order to compare with previous Pol II-ChIP data for this tissue [PMID 22821985].

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Cell-type specific and combinatorial usage of diverse transcription factors revealed by genome-wide binding studies in multiple human cells [new ChIP-Seq samples]

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