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Series GSE95121 Query DataSets for GSE95121
Status Public on May 21, 2018
Title Priming and Activation of ERα Enhancers Through Ordered and Cooperative Interactions Between p300 and Mediator [ChIP-seq]
Organism Homo sapiens
Experiment type Genome binding/occupancy profiling by high throughput sequencing
Summary Estrogen receptors alpha (ERα) converges estrogen signaling by orchestrating estrogen (E2)-dependent transcription regulation. Upon binding to the chromatin, ERα serves as a nucleation site for de novo formation of transcription enhancer complexes. Steroid receptor coactivators family proteins (SRCs, a.k.a p160) and Mediator complex are the two coregulators for ERα that directly interact with the receptors and control enhancer activity by regulating recruitment of other coregulators and transcription machinery. Lysine acetyltransferase p300/CBP is also a critical coregulator for ERα that is recruited through SRCs in stable ERα enhancer complex. ERα enhancers exhibit common enhancer features including enrichment of enhancer-specific histone modification (e.g. H3K4me1 and H3K27ac), coregulator recruitment, and active transcription. Despite of recognition of those enhancer characteristics, the order of assembly and functions and the functional relationships among enhancer features during active enhancer complex formation is not well understood. Using the time course genomics and molecular biology assays, we dissect the order of assembly and functions of ERα enhancer complex formation. We describe the mechanism of SRC-independent p300 recruitment mediated by Mediator, leading to initial partial activation of the ERα enhancer for enhance priming. Maturation of the enhancer in turn switches the p300 recruitment mechanism to an SRC-dependent manner, resulting in the maximum enhancer activity and transcription outcomes. Thus, our results illustrate the role of SRC in enhancing the activity of primed enhancers. Furthermore, we show that forced recruitment of p300 to inactive enhancers establishes active enhancer marks and induces ERα-target gene expression. Together, we demonstrate the molecular mechanisms of ERα enhancer complex formation where p300 plays a pivotal role in enhancer activation and target gene expression controlled by distinctive mechanisms through different stages of ERα enhancer complex formation.
 
Overall design ChIP-seq (ERα, Med1, SRC, p300, and H3K27ac) and GRO-seq datasets were generated using E2-treated MDA-MB-231 cells expressing ERα wt and L540Q mutant to define ERα enhancers and to determine enhancer complex formation and activity.

ChIP antibody information;
ERa, rabbit polyclonal antiserum generated in the Kraus Laboratory against the 1-113 of human ERa
Med1, Cat. # A300-793A, Lot # A300-793A-2
SRC, rabbit polyclonal antiserum generated in the Kraus Laboratory against the 624-1130 of mouse SRC2
H3K27ac, Cat. # ab4729, Lot # GR254711-1
p300, Cat. # sc-584x, Lot # I1715
 
Contributor(s) Murakami S, Nagari A, Kraus WL
Citation(s) 28887413
Submission date Feb 21, 2017
Last update date May 15, 2019
Contact name W. Lee Kraus
E-mail(s) lee.kraus@utsouthwestern.edu
Organization name UT Southwestern Medical Center
Street address 5323 Harry Hines Blvd.
City Dallas
State/province TX
ZIP/Postal code 75390-8511
Country USA
 
Platforms (2)
GPL11154 Illumina HiSeq 2000 (Homo sapiens)
GPL18573 Illumina NextSeq 500 (Homo sapiens)
Samples (79)
GSM2496463 Input_for_ERα_Med1_H3K27ac_SRC_ChIP-seq_231ERαWT_0min_rep1
GSM2496464 Input_for_ERα_Med1_H3K27ac_SRC_ChIP-seq_231ERαWT_0min_rep2
GSM2496465 Input_for_ERα_Med1_H3K27ac_SRC_ChIP-seq_231ERαWT_0min_rep3
This SubSeries is part of SuperSeries:
GSE95123 Priming and Activation of ERα Enhancers Through Ordered and Cooperative Interactions Between p300 and Mediator
Relations
BioProject PRJNA376133
SRA SRP100436

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE95121_ERalpha_LQ_0m.bw 60.8 Mb (ftp)(http) BW
GSE95121_ERalpha_LQ_45m.bw 54.9 Mb (ftp)(http) BW
GSE95121_ERalpha_WT_0m.bw 61.3 Mb (ftp)(http) BW
GSE95121_ERalpha_WT_45m.bw 61.0 Mb (ftp)(http) BW
GSE95121_H3K27Ac_LQ_0m.bw 62.2 Mb (ftp)(http) BW
GSE95121_H3K27Ac_LQ_45m.bw 62.6 Mb (ftp)(http) BW
GSE95121_H3K27Ac_WT_0m.bw 63.9 Mb (ftp)(http) BW
GSE95121_H3K27Ac_WT_45m.bw 66.5 Mb (ftp)(http) BW
GSE95121_Input_LQ_0m.bw 66.4 Mb (ftp)(http) BW
GSE95121_Input_LQ_45m.bw 64.6 Mb (ftp)(http) BW
GSE95121_Input_WT_0m.bw 68.9 Mb (ftp)(http) BW
GSE95121_Input_WT_45m.bw 62.5 Mb (ftp)(http) BW
GSE95121_Med1_LQ_0m.bw 69.5 Mb (ftp)(http) BW
GSE95121_Med1_LQ_45m.bw 71.5 Mb (ftp)(http) BW
GSE95121_Med1_WT_0m.bw 70.5 Mb (ftp)(http) BW
GSE95121_Med1_WT_45m.bw 71.6 Mb (ftp)(http) BW
GSE95121_SRC_LQ_0m.bw 70.2 Mb (ftp)(http) BW
GSE95121_SRC_LQ_45m.bw 71.7 Mb (ftp)(http) BW
GSE95121_SRC_WT_0m.bw 58.4 Mb (ftp)(http) BW
GSE95121_SRC_WT_45m.bw 72.8 Mb (ftp)(http) BW
GSE95121_p300_LQ_0m.bw 64.4 Mb (ftp)(http) BW
GSE95121_p300_LQ_20m.bw 99.5 Mb (ftp)(http) BW
GSE95121_p300_LQ_45m.bw 63.5 Mb (ftp)(http) BW
GSE95121_p300_WT_0m.bw 69.3 Mb (ftp)(http) BW
GSE95121_p300_WT_20m.bw 101.8 Mb (ftp)(http) BW
GSE95121_p300_WT_45m.bw 69.6 Mb (ftp)(http) BW
GSE95121_p300input_LQ_20m.bw 61.8 Mb (ftp)(http) BW
GSE95121_p300input_WT_20m.bw 61.8 Mb (ftp)(http) BW
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