Abstract
Bromodomain-containing protein 4 (BRD4) is an important epigenetic reader implicated in the pathogenesis of a number of different cancers and other diseases. Brd4-null mouse embryos die shortly after implantation and are compromised in their ability to maintain the inner cell mass, which gives rise to embryonic stem cells (ESCs). Here we report that BRD4 regulates expression of the pluripotency factor Nanog in mouse ESCs and preimplantation embryos, as well as in human ESCs and embryonic cancer stem cells. Inhibition of BRD4 function using a chemical inhibitor, small interfering RNAs, or a dominant-negative approach suppresses Nanog expression, and abolishes the self-renewal ability of ESCs. We also find that BRD4 associates with BRG1 (brahma-related gene 1, aka Smarca4 (SWI/SNF-related, matrix-associated, actin-dependent regulator of chromatin, subfamily a, member 4)), a key regulator of ESC self-renewal and pluripotency, in the Nanog regulatory regions to regulate Nanog expression. Our study identifies Nanog as a novel BRD4 target gene, providing new insights for the biological function of BRD4 in stem cells and mouse embryos. Knowledge gained from these non-cancerous systems will facilitate future investigations of how Brd4 dysfunction leads to cancers.
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Abbreviations
- Brd4:
-
bromodomain-containing protein 4
- ICM:
-
inner cell mass
- ESCs:
-
embryonic stem cells
- BET:
-
bromodomain and extraterminal protein
- LIF:
-
leukemia inhibitory factor
- BMP4:
-
bone morphogenetic protein 4
- AP:
-
alkaline phosphatase
- siRNAs:
-
small interfering RNAs
- BDI/II:
-
double bromodomains
- ChIP:
-
chromatin immunoprecipitation
- BRG1:
-
brahma-related gene 1, aka Smarca4 (SWI/SNF-related, matrix-associated, actin-dependent regulator of chromatin, subfamily a, member 4)
- ChIP-reChIP:
-
sequential ChIP analysis
- H3K27me3:
-
trimethylation of Lys27 of histone 3
- FBS:
-
fetal calf serum
- GV:
-
germinal vesicle
- MII:
-
metaphase II
- PVP:
-
polyvinylpyrrolidone
- KSOM+AA:
-
KSOM medium containing amino acids
- RA:
-
retinoic acid
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Acknowledgements
We thank M Celeste Simon (University of Pennsylvania) for P19 cells, Mitchell J Weiss (The Children’s Hospital of Philadelphia) for R1 cells, James E Bradner (Dana-Farber Cancer Institute) for JQ1, Weidong Wang (NIH) for Brg1 antibody and Jing Jiao for technical support. We thank members of our laboratories for helpful discussion and critical review of the manuscript. This work has been supported by NIH grants R01CA148768 and R01CA142723 to JY and R01HD022681 to RMS.
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Liu, W., Stein, P., Cheng, X. et al. BRD4 regulates Nanog expression in mouse embryonic stem cells and preimplantation embryos. Cell Death Differ 21, 1950–1960 (2014). https://doi.org/10.1038/cdd.2014.124
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DOI: https://doi.org/10.1038/cdd.2014.124
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