Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Detection of transcription factor NRF1, YY1 and SP1 bound regions in human peripheral blood monocytes

ABSTRACT: To study the correlation between sequence motif appearance, transcription factor binding and aberrant hypermethylation in the cell lines, we performed ChIP-on-chip analyses (on CpG island microarrays) for the transcription factors Sp1, NRF1 and YY1 in normal peripheral blood monocytes. Keywords: ChIP-on-Chip; comparative genomic hybridization Transcription factor bound genomic DNA was enriched using ChIP. On each microarray, the enriched material was compared to the genomic input to identify transcription factor bound regions. Two biological replicates were analysed for each factor.

ORGANISM(S): Homo sapiens

SUBMITTER: Christian Schmidl

PROVIDER: E-GEOD-16078 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

General transcription factor binding at CpG islands in normal cells correlates with resistance to de novo DNA methylation in cancer cells.

Gebhard Claudia C Benner Chris C Ehrich Mathias M Schwarzfischer Lucia L Schilling Elmar E Klug Maja M Dietmaier Wolfgang W Thiede Christian C Holler Ernst E Andreesen Reinhard R Rehli Michael M

Cancer research 20100209 4

Aberrant DNA methylation at CpG islands is thought to contribute to cancer initiation and progression, but mechanisms that establish and maintain DNA methylation status during tumorigenesis or normal development remain poorly understood. In this study, we used methyl-CpG immunoprecipitation to generate comparative DNA methylation profiles of healthy and malignant cells (acute leukemia and colorectal carcinoma) for human CpG islands across the genome. While searching for sequence patterns that ch ...[more]

PMID: 20145141

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Project description:We have previously developed an approach that fractionates genomic DNA fragments depending on their CpG density (methyl-CpG-immunoprecipitation, MCIp), and adapted this approach to identify regions that are differentially methylated in the two closely related regulatory T-cells (Treg cells) and conventional T-cells (Tconv cells). Because Treg cells naturally occur at a relatively low frequency, we used a previously established protocol to expand Treg cells from a stable naÃ¯ve Treg population that is characterized by the co-expression of CD4, CD25 and CD45RA. We separated gDNA of both expanded T cell lineages (Tregexp and Tconvexp) into unmethylated (CpG) and methylated pools (mCpG) using MCIp and compared cell type-specific differences in DNA methylation by co-hybridization of the two umethylated or the two methylated DNA subpopulations of Treg and Tconv, respectively, to these locus-wide custom tiling arrays. As enriched DNA-fragments from a cell type in the methylated fraction should be depleted in the unmethylated fraction, the signal intensities in CpG pool and mCpG pool hybridizations should complement themselves (âMirror-Imageâ approach) and thereby allow the identification of differentially methylated regions (DMR). Because we expected to find lineage-specific methylation differences with greater probability in regions associated with differential transcriptional activity, we limited our analysis to gene loci that showed cell type-specific gene expression in Treg versus Tconv cells plus a handful of control regions that were equally expressed in both cell types. The microarray used in this study covered 12 megabases of the human genome and contained 69 regions (with a median size of 100.000 kb) and 128 proximal promoter regions and 181 genes, which included a number of well known and functionally relevant genes like CD40LG, IFNG, FOXP3, IL2RA and CTLA4. Keywords: MCIp-on-chip; comparative genomic hybridization With MCIp gDNA from Treg or Tconv cells was separated into hypo- and hypermethylated pools. On each array, wheather the two hypomethylated fractions- one from Treg, the other from Tconv cells- or the two hypermethylated fractions were cohybridized. Two biological replicates.

Dataset Information

Detection of transcription factor NRF1, YY1 and SP1 bound regions in human peripheral blood monocytes

Publications

General transcription factor binding at CpG islands in normal cells correlates with resistance to de novo DNA methylation in cancer cells.

Similar Datasets

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