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FOXP3 binding sites in promoter regions of human Jurkat T cells

ABSTRACT: The transcription factor (TF) Forkhead Box P3 (FOXP3) is constitutively expressed in high levels in natural occurring CD4+CD25+ regulatory T cells (nTreg) and is not only the most accepted marker for that cell population, but is considered lineage determinative. Chromatin immunoprecipitation (ChIP) of transcription factors in combination with genomic tiling microarray analysis (ChIP-on-Chip) has been shown to be an appropriate tool to identify FOXP3 transcription factor binding sites (TFBS) on a genome-wide scale. In combination with microarray expression analysis the ChIP-on-Chip technique allows to identify direct FOXP3 target genes. ChIP-on-Chip analysis of human FOXP3Δ2 isoform expressed in resting and PMA / ionomycin stimulated Jurkat T cells revealed several thousand putative FOXP3 binding sites and importance of intronic regions for FOXP3 binding. Knowledge of general distribution patterns of FOXP3 TFBS in the human genome under resting and activated conditions contributes to a better understanding of this TF and its influence on direct target genes with importance for Treg cell phenotype and function.

ORGANISM(S): Homo sapiens

PROVIDER: GSE37252 | GEO | 2012/11/26

SECONDARY ACCESSION(S): PRJNA159257

REPOSITORIES: GEO

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FOXP3 binding sites in promoter regions of human Jurkat T cells

Project description:The transcription factor (TF) Forkhead Box P3 (FOXP3) is constitutively expressed in high levels in natural occurring CD4+CD25+ regulatory T cells (nTreg) and is not only the most accepted marker for that cell population, but is considered lineage determinative. Chromatin immunoprecipitation (ChIP) of transcription factors in combination with genomic tiling microarray analysis (ChIP-on-Chip) has been shown to be an appropriate tool to identify FOXP3 transcription factor binding sites (TFBS) on a genome-wide scale. In combination with microarray expression analysis the ChIP-on-Chip technique allows to identify direct FOXP3 target genes. ChIP-on-Chip analysis of human FOXP3M-NM-^T2 isoform expressed in resting and PMA / ionomycin stimulated Jurkat T cells revealed several thousand putative FOXP3 binding sites and importance of intronic regions for FOXP3 binding. Knowledge of general distribution patterns of FOXP3 TFBS in the human genome under resting and activated conditions contributes to a better understanding of this TF and its influence on direct target genes with importance for Treg cell phenotype and function. ChIP-DNA from FOXP3(M-NM-^T2) expressing Jurkat T cells under resting and PMA / ionomycin stimulated conditions from duplicate experiments was analyzed. FOXP3-specific tiling array data were analyzed in reference to an individual isotype control dataset (J-FOXP3 ChIP'd with FOXP3 antibody vs. J-FOXP3 ChIP'd with isotype control antibody). In total 8 tiling array analyses were performed (2x resting J-FOXP3 with FOXP3-IP, 2x resting J-FOXP3 with isotype-IP, 2x PMA/iono J-FOXP3 with FOXP3-IP, 2x PMA/iono J-FOXP3 with isotype-IP)

2012-11-26 | E-GEOD-37252 | biostudies-arrayexpress

Identification of FOXP3-dependent transcripts in human FOXP3 expressing Jurkat T cells

2012-11-26 | E-GEOD-37253 | biostudies-arrayexpress

Identification of FOXP3-dependent transcripts in human FOXP3 expressing Jurkat T cells

2012-11-26 | GSE37253 | GEO

Genome wide FOXP3 binding sites in human cord blood derived CD4+CD25+ natural T regulatory cells (nTreg)

Project description:FOXP3 is essential for the stability and function of nTReg. We have characterised the FOXP3-dependent cis-regulatory network through a combination of whole genome ChIP-on-chip and transcriptional profiling of primary human nTreg cells . Human nTreg cells were isolated from cord blood and expanded ex vivo (day9). These analyses identified approximately 8,000 high quality FOXP3 binding sites the majority (85%) of which were located in proximity to annotated genes . Intersection of DNA binding data with gene expression profiles predicted 739 candidate target genes. The supplementary bed file contains all 8305 high quality binding FOXP3 binding regions reported in the paper.

2010-06-22 | GSE20995 | GEO

DNA methylation differences between human regulatory T cells and conventional T cells

Project description:Regulatory T cells (Treg) prevent the emergence of autoimmune disease. Prototypic natural Treg (nTreg) are programmed by Forkhead-box P3 (FOXP3) and can be reliably identified by demethylation at the FOXP3 locus. To explore the nTreg methylation landscape we performed genome-wide methylation studies on human naïve resting nTreg (rTreg) and conventional naïve CD4+ T cells (Naïve). We detected 2,315 differentially methylated CpGs between these two cell types, many of which clustered into 127 regions of differential methylation (RDMs). T cell activation induced changes in 466 individual CpGs and 18 RDMs in naïve CD4+ T cells, but did not alter DNA methylation in rTreg. Expression of mRNA for TIGIT, an immune suppressive receptor demethylated in rTreg, was upregulated in nTreg and reduced in peripheral blood mononuclear cells of individuals at risk for autoimmune (type 1) diabetes. Gene-set testing of the 127 RDMs revealed enrichment of common Treg signature genes, FOXP3 bound genes and genes directly upregulated by FOXP3, which was primarily driven by the subset of demethylated RDMs. A putative Forkhead-binding motif overrepresented in promoter-associated RDMs suggests methylation regulates gene expression by influencing FOXP3 binding. Our findings provide new insights into epigenetic regulation of human nTreg and the potential to exploit differential methylation as an immune biomarker in human diseases.

2013-09-09 | GSE49667 | GEO

DNA methylation differences between human regulatory T cells and conventional T cells

2013-09-09 | E-GEOD-49667 | biostudies-arrayexpress

Transcription Factor Binding Sites clustered Regions of 133 Cell Lines

Project description:Purpose: 224 GSM Samples form GSE32970 and GSE29692 was reanalyzed to find the TFBS-clustered regions of 133 cell lines. TFBS-clustered regions were divided into ten classes belong to the TF complexity. Methods: 1. We assigned the binding sites of 542 TFs in 133 cell lines as our record GSE53962 . 2. We performed a Gaussian kernel density estimation across the genome with a bandwidth of 300 bp, using the centers of each of the TF binding peaks as points. Then, we scanned this density for peaks, and denoted each peak a TF region.To determine the complexity of the TF region, we summed the Gaussian kernalized distance from the peak to each TF that contributed at least 0.1 to its strength. The TF region around eat peak was derived by finding the maximum distance (in bp) from the peak to a contributing TF, and then adding 150 bp (one half of the bandwidth). Each TF region is centered on the peak, and have a TF complexity value. 3. According to TFBS complexity, we divided these TFBS-clustered regions into ten classes: from TC0 to TC9 with increasing TFBS complexity. Result: Using the binding sites of 542 TFs in 133 cell lines, we assigned a TF complexity score to each TF region corresponding to the number of distinct TFs bound, resulting in ten classes TFBS-clustered regions of 133 cell lines.

2014-10-03 | GSE59016 | GEO

Bacterial transcription factors binding to coding regions regulate the internal cryptic promoters

Project description:To analyze the antisense transcripts, we performed LM-seq for Pesudomonas aeruginosa ∆algR and Pesudomonas syringae ∆rhpS and their corresponding WT strain. Date-minging in ChIP-seq and HT-SELEX results from public dataset, we got the TF binding sites (TFBS) across genome. Coupling analysis with LM-seq and these TFBS, our results revealed that the coding region binding TFs regulate antisense RNA transcription by regulating the activity of cryptic promoters.

2022-09-28 | GSE175852 | GEO

Gene expression profiling of in vivo derived induced and natural FOXP3+ regulatory T cells in the mouse

Project description:The relative contribution of induced and natural Foxp3+ regulatory T cells (iTreg and nTreg cells, respectively) to the maintenance of tolerance is unknown. We examined their respective roles by in vivo adoptive transfer immunotherapy of newborn Foxp3-deficient BALB/c mice. Survival, weight gain, tissue infiltration, T cell activation, and the concentration of proinflammatory cytokines were used as outcome measurements. Treatment with iTreg cells alone was not successful. While effective in preventing death, treatment with nTreg cells alone was associated with chronic inflammation and autoimmunity. Outcomes markedly improved when conventional T (Tconv) cells were transferred together with the nTreg cells, where 10% of the peripheral Treg cell pool was derived by in-situ conversion. This enhancement depended upon the capacity of Tconv cells to express Foxp3. The gene expression profile of in vivo derived iTreg cells was similar to the established nTreg cell genetic signature. These results identify iTreg cells as an essential regulatory subset that supplements tolerance maintained by nTreg cells.

2011-06-30 | GSE19512 | GEO

The DNA affinity purification DAP-Seq assay [ChIP-seq]

Project description:As 5-15% of higher eukaryotes genes are transcription factors (TFs), the lack of transcription factor binding site (TFBS) information for most factors in most organisms limits the study of gene regulation. Here we describe a next-generation sequencing method, DNA affinity purification (DAP-Seq), an in vitro gDNA/TF interaction assay that produces whole-genome TFBS annotation for any factor from any organism. Like ChIP-Seq, DAP-Seq resolves TFBS as discrete peaks at genomic locations which allows for accurate motif prediction direct assignment of functionally relevant target genes, and shows better overlap with ChIP-Seq peaks than indirect motif assignment approaches. We applied DAP-Seq to a set of 50 transcription factors in eight Arabidopsis thaliana and one Zea Mays families to gain novel biological insight into TFBS architectures, functions, evolution and methylation-sensitivity. Overall, DAP-Seq offers a low-cost high-throughput approach to identify TFBS in native sequence context for any organism complete with all DNA chemical modifications.

2016-05-25 | GSE60142 | GEO

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