Enhancer ID: | E_01_239 | |
Enhancer symbol: | -- | |
Species: | Human | |
Position : | chr1:181117806-181122451 |
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Biosample name: | HCT 116 | |
Experiment class : | Low+High throughput |
Enhancer type: | Enhancer | |
Disease: | Cervical Cancer | |
DO: | DOID:4362 | |
Mesh: | D002583 | |
Distance from TSS: | >2KB | |
Pubmed ID: | 26754925 |
Enhancer experiment: | ChIP-seq,Luciferase Reporter Assay,4C-seq | |
Enhancer experiment description: | ChIP-seq analysis identified two p53 bind-ing sites 11 kb (RE1) and 46 kb (RE2) downstream of the IER5 gene (Fig. 1F).p53 binding to RE2 was strong,while binding to RE1 was relatively weak. In addition, H3K27 acetylation was detected at RE2, and was strongly increased by 5-FU treatment, showing that RE2 is an active enhancer that is highly activated upon DNA damage.We found sequences highly similar to the p53 consensus binding sequence at the center of RE2 (TRANSFAC match score; 0.94) but not in RE1 (TRANSFAC match score; 0.74, Fig. 1G). We generated DNA oligonucleotides representing RE1 and RE2, and created a version of RE2 mutated in the p53 consensus sequence (RE2 mut; sequences shown in Fig. 1G), and cloned each upstream of a luciferase reporter gene containing a minimal promoter. As shown in Fig. 1H, p53 strongly activated a reporter containing RE2, but not RE2 mut or RE1. We further performed 4C-seq analysis to identify chromatin regions that might interact with RE2. As shown in Fig.1F, we detected chromatin interaction between RE2 and RE1, and between RE2 and the IER5 gene promoter region, in both HCT116 p53 + /+ and −/− cells. The interactions between the RE2 and the IER5 promoter region seem to be stabilized by 5-FU treatment in HCT116 p53 +/+ cells but not in HCT116 p53 −/− cells. Collectively, these results show that RE2 is localized in the vicinity of the IER5 promoter when a higher order chromatin structure is formed, and identifies RE2 as the p53-responsive element of the IER5 gene. |
Target gene : | IER5(SBBI48) | |
Strong evidence: | 4C-seq | |
Less strong evidence: | Luciferase Reporter Assay | |
Target gene experiment description: | We generated DNA oligonucleotides representing RE1 and RE2, and created a version of RE2 mutated in the p53 consensus sequence (RE2 mut; sequences shown in Fig. 1G), and cloned each upstream of a luciferase reporter gene containing a minimal promoter. As shown in Fig. 1H, p53 strongly activated a reporter containing RE2, but not RE2 mut or RE1. We further performed 4C-seq analysis to identify chromatin regions that might interact with RE2. As shown in Fig.1F, we detected chromatin interaction between RE2 and RE1, and between RE2 and the IER5 gene promoter region, in both HCT116 p53 + /+ and −/− cells. The interactions between the RE2 and the IER5 promoter region seem to be stabilized by 5-FU treatment in HCT116 p53 +/+ cells but not in HCT116 p53 −/− cells. Collectively, these results show that RE2 is localized in the vicinity of the IER5 promoter when a higher order chromatin structure is formed, and identifies RE2 as the p53-responsive element of the IER5 gene. |
TF name : | TP53(BCC7,BMFS5,LFS1,P53,TRP53) | |
TF experiment: | Luciferase Reporter Assay | |
TF experiment description: | ChIP-seq analyses were performed using antibodies against p53, H3K27ac, H3K4me1, H3K4me3 and phospho-RNAP II. Two p53 binding sites RE1 (11 kb downstream) and RE2 (46 kb downstream) were identified. |
Enhancer function : | -- |
Enhancer function experiment: | -- |
Enhancer function experiment description: |
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SNP ID: | -- | |
SNP position: | -- |
SNP experiment: | -- |
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