Obesity accounts for a substantial and growing burden of global disease, leading to high morbidity and mortality in type 2 diabetes, cardiovascular disease, and certain cancers. Brown adipose tissue (BAT) dissipates energy as heat and has been proposed as a therapeutic target to combat obesity. The unique thermogenic capacity of BAT is primarily conferred by uncoupling protein 1 (Ucp1) , which dissipates energy by uncoupling the proton motive force from ATP generation. To identify novel transcriptional regulators of Ucp1, we performed an unbiased CRISPR/Cas9-based chromatin immunoprecipitation assay (enChIP) and identified H2A.Z as a novel trans-acting factor recruited to the promoter and enhancer of Ucp1 by β3-adrenergic receptor (β3-AR) stimulation. H2A.Z is an essential and evolutionary conserved histone variant of the canonical histone H2A. Knockdown of H2A.Z in brown adipocytes led to lower β3-AR-induced Ucp1 level and impeded the expression of other known thermogenic genes (e.g., Cidea, Pgc1a) , leading to impaired cellular thermogenesis. Mechanistically, by profiling the genome-wide distribution of H2A.Z via ChIP-seq, we observed H2A.Z was enriched at promoter and enhancer of Ucp1 upon β3-AR stimulation, which is consistent with the enChIP result. Intriguingly, we found that the average occupancy of H2A.Z in the promoter and enhancer regions of other β3-AR-upregulated genes (e.g., Zfp516, Vdr, Pck1, Gys2) was also higher than that in non-stimulated condition, indicating a global regulatory role of H2A.Z in the thermogenic program in response to β3-AR stimulation. Additionally, in the enhancer regions of β3-AR-induced genes, we also detected an increased level of H2A.Z acetylation, which is a mark of active chromatin and confers an open chromatin conformation. Taken together, these findings reveal the essential role of H2A.Z-mediated chromatin organization in BAT thermogenic function.
Y.Zhang: None. R.Zheng: None. C.Wang: None. J.Darcy: None. K.Chen: None. Y.Tseng: Consultant; Cellarity.
National Institutes of Health (R01DK102898)