 |
 |
GEO help: Mouse over screen elements for information. |
|
| Status |
Public on Jan 12, 2021 |
| Title |
BACH1 Recruits NANOG and Histone H3 Lysine 4 Methyltransferase MLL/SET1 Complexes to Regulate Enhancer-promoter Activity and Maintains Pluripotency |
| Organism |
Mus musculus |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
Expression profiling by high throughput sequencing
|
| Summary |
Maintenance of stem-cell identity requires proper regulation of enhancer activity. Both transcription factors OCT4/SOX2/NANOG and histone methyltransferase complexes MLL/SET1 were shown to regulate enhancer activity, but how they are regulated in embryonic stem cells (ESCs) remains further studies. Here, we report a transcription factor BACH1, who which directly interacts with OCT4/SOX2/NANOG (OSN) and MLL/SET1 methyltransferase complexes and maintains pluripotency in mouse ESCs (mESCs). BTB domain and bZIP domain of BACH1 are required for these interactions and pluripotency maintenance. Loss of BACH1 reduced the interaction between NANOG and MLL1/SET1 complexes, and decreased their occupancy on chromatin, and further decreased H3 lysine 4 trimethylation (H3K4me3) level on gene promoters and (super-) enhancers, leading to decreased enhancer activity and transcription activity, especially on stemness-related genes. Moreover, BACH1 recruited NANOG through chromatin looping and regulated remote NANOG binding, fine-tuning enhancer-promoter activity and gene expression. Collectively, these observations suggest that BACH1 maintains pluripotency in ESCs by recruiting NANOG and MLL/SET1 complexes to chromatin and maintaining the trimethylated state of H3K4 and enhancer-promoter activity, especially on stemness-related genes.
|
| |
|
| Overall design |
Bach1 knock-out (Bach1-KO) mESCs were generated by CRISPR/Cas9. DoxBach1 mESCs expressing dox inducible Bach1 with C-terminal Flag tag were generated by piggyBac (PB) transposon system. We performed ChIP-seq of Nanog, H3K4me1, H3K4me3, and H3K27ac in wild-type (WT) mESCs and Bach1-KO mESCs, and also the ChIP-seq of Flag tagged Bach1 in DoxBach1 mESCs. We also perfromed the RNA-seq of WT mESCs and Bach1-KO mESCs. RNA-seq for each cell line had biological triplicates.
|
| |
|
| Contributor(s) |
Meng D, Wang S |
| Citation(s) |
33503260 |
| |
| Submission date |
Dec 23, 2019 |
| Last update date |
Mar 10, 2021 |
| Contact name |
Siqing Wang |
| E-mail(s) |
wangs16@chop.edu
|
| Organization name |
Children’s Hospital of Philadelphia
|
| Department |
Department of Pediatrics, Division of Hematology
|
| Lab |
Gerd Blobel's Lab
|
| Street address |
3401 Civic Center Blvd
|
| City |
Philadelphia |
| State/province |
PENNSYLVANIA |
| ZIP/Postal code |
19104 |
| Country |
USA |
| |
|
| Platforms (2) |
| GPL21273 |
HiSeq X Ten (Mus musculus) |
| GPL24247 |
Illumina NovaSeq 6000 (Mus musculus) |
|
| Samples (23)
|
|
| Relations |
| BioProject |
PRJNA597299 |
| SRA |
SRP238575 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE142519_RAW.tar |
4.5 Mb |
(http)(custom) |
TAR (of BED, BROADPEAK, NARROWPEAK) |
| GSE142519_RNASEQ_TPM_2020.xlsx |
2.2 Mb |
(ftp)(http) |
XLSX |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
| Processed data are available on Series record |
|
|
|
|
 |
Less...
More...