Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

POLII localization in HeLa cells

ABSTRACT: RNA polymerase II (POLR2A; POLII) was localized via ChIP-chip using N20 Ab-mediated pull-down of POLII Classical design; ChIP-chip of POLII in ENCODE regions

ORGANISM(S): Homo sapiens

SUBMITTER: Mark Bieda

PROVIDER: E-GEOD-4337 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Unbiased location analysis of E2F1-binding sites suggests a widespread role for E2F1 in the human genome.

Bieda Mark M Xu Xiaoqin X Singer Michael A MA Green Roland R Farnham Peggy J PJ

Genome research 20060410 5

The E2F family of transcription factors regulates basic cellular processes. Here, we take an unbiased approach towards identifying E2F1 target genes by examining localization of E2F1-binding sites using high-density oligonucleotide tiling arrays. To begin, we developed a statistically-based methodology for analysis of ChIP-chip data obtained from arrays that represent 30 Mb of the human genome. Using this methodology, we identified regions bound by E2F1, MYC, and RNA Polymerase II (POLR2A). We f ...[more]

PMID: 16606705

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Project description:Recent genome-scale ChIP-chip studies of transcription factors have shown that a low percentage of experimentally determined binding sites contain the consensus motif for the immunoprecipitated factor. In most cases, differences between in vivo target sites that contain or lack a consensus motif have not been explored. We have previously shown that most sites to which E2F family members are bound in vivo do not contain E2F consensus motifs. The main purpose of this study was to develop an understanding of how E2F binding specificity is achieved in vivo. In particular, we have addressed how E2F family members are recruited to core promoter regions that lack a consensus motif and are excluded from other regions that contain a consensus motif. Using promoter and ENCODE arrays, we have shown that the predominant factors specifying whether E2F is recruited to an in vivo binding site are a) the site must be in a core promoter and b) the promoter region must be utilized as a promoter by the transcriptional machinery in that particular cell type. We have tested three models for recruitment of E2F to core promoters lacking a consensus site, including a) indirect recruitment, b) looping to the core promoter mediated by an E2F bound to a distal consensus motif, and c) assisted binding of E2F to a site that weakly resembles an E2F consensus motif within the core promoter. To test these models, we developed a new in vivo assay, termed eChIP, which allows analysis of transcription factor binding to isolated promoter fragments. Our findings suggest that in vivo a) the presence of a consensus motif is not sufficient to recruit E2Fs, b) E2Fs can bind to isolated regions that lack a consensus motif, and c) binding can require regions other than the best match to the E2F PWM in the core promoter. Keywords: E2F, ChIP-chip, transcription factor binding, consensus motifs 37 ChIP-chip arrays (of these, 14 array sets are biological duplicates). 22 samples are included in this series, the rest can be found in supplementary info to the following papers: Xu 2007, Jin 2006, Komashko 2008

Dataset Information

POLII localization in HeLa cells

Publications

Unbiased location analysis of E2F1-binding sites suggests a widespread role for E2F1 in the human genome.

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