Brown adipose tissue (BAT) dissipates chemical energy in the form of heat as a defence against hypothermia and obesity. Current evidence indicates that brown adipocytes arise from Myf5+ dermotomal precursors through the action of PR domain containing protein 16 (PRDM16) transcriptional complex1,2. However, the enzymatic component of the molecular switch that determines lineage specification of brown adipocytes remains unknown. Here we show that euchromatic histone-lysine N-methyltransferase 1 (EHMT1) is an essential BAT-enriched lysine methyltransferase in the PRDM16 transcriptional complex and controls brown adipose cell fate. Loss of EHMT1 in brown adipocytes causes a severe loss of brown fat characteristics and induces muscle differentiation in vivo through demethylation of histone 3 lysine 9 (H3K9me2 and 3) of the muscle-selective gene promoters. Conversely, EHMT1 expression positively regulates the BAT-selective thermogenic program by stabilizing the PRDM16 protein. Notably, adipose-specific deletion of EHMT1 leads to a marked reduction of BAT-mediated adaptive thermogenesis, obesity and systemic insulin resistance. These data indicate that EHMT1 is an essential enzymatic switch that controls brown adipose cell fate and energy homeostasis.
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We are grateful to A. Tarakhovsky, E. D. Rosen, Y. Shinkai and E. Hara for providing mice and plasmids. We thank our colleagues in the University of California, San Francisco, including Y. Qiu, A. Chawla, C. Paillart, S. Koliwad, M. Robblee, D. Scheel, S. Ohata, L. Mera, D. Lowe, S. Sonne, S. Keylin, I. Luijten, H. Hong and E. Tomoda for their assistance. This work was supported by grants from the National Institutes of Health (DK087853 and DK97441) to S.K. We acknowledge supports from the DERC center grant (DK63720), University of California, San Francisco Program for Breakthrough Biomedical Research program, the Pew Charitable Trust, and PRESTO from the Japan Science and Technology Agency to S.K. H.O. is supported by the Manpei Suzuki Diabetes Foundation. K.S. is supported by a fellowship from the Japan Society for the Promotion of Science.
The authors declare no competing financial interests.
Extended data figures and tables
a, The putative BAT was micro-dissected from WT and Ehmt1myf5 knockout embryos. mRNA expression of Prdm16 was measured by qRT–PCR. Data are presented as mean and s.e.m. (n = 8–10). b, Western blotting to detect endogenous EHMT1, PRDM16, UCP1 and MHC in BAT from WT and Ehmt1myf5 knockout embryos. α-Tublin protein was shown as a loading control.
Extended Data Figure 2 Ectopic activation of skeletal-muscle-selective genes and reduction of BAT-selective genes in the BAT from Ehmt1adipo knockout mice.
a, Western blotting for endogenous EHMT1 in BAT and liver from WT and Ehmt1adipo knockout mice. β-Actin protein was shown as a loading control. b, Amounts of mRNA expression of BAT, skeletal muscle, white fat and beige-fat selective genes in BAT from Ehmt1adipo knockout mice. Values were normalized to those in WT mice. The amounts of mRNA were visualized by a heat-map using Multi Experiment Viewer. c, Venn diagram showing the overlapped genes between Ehmt1myf5 knockout and Ehmt1adipo knockout mice. RNA-sequencing and gene ontology analyses identified 33 genes that were similarly dysregulated both in the Ehmt1myf5 knockout BAT and the Ehmt1adipo knockout BAT. The mRNA expression values were normalized to WT mice for each knockout model and visualized by a heat-map using Multi Experiment Viewer. The colour scale shows the amounts of mRNA of the genes in a blue (low)–white (no change)–red (high) scheme.
a, The b3-AR agonist CL316,243 at a dose of 0.5 mg kg−1 or saline were administered to WT or Ehmt1adipo knockout mice for 7 days. Inguinal WAT was collected for gene expression analysis. Amounts of mRNA expression of BAT and beige-fat selective genes (as indicated) were measured by qRT–PCR (n = 3–6). †Significant between saline and CL316,243 in WT mice. b, Immunohistochemistry for UCP1 in a. Scale bar, 100 μm. Nuclei were stained with DAPI. c, To test a cell-autonomous requirement for EHMT1 in beige/brite cell development, the stromal vascular (SV) fractions were isolated from the inguinal WAT of Ehmt1flox/flox mice. Cells were infected with adenovirus expressing GFP or Cre. The SV cells were differentiated in the presence or absence of rosiglitazone (Rosi) at 0.5 μM. Amounts of mRNA expression of BAT-selective genes (as indicated) were measured by qRT–PCR. Deletion of Ehmt1 was confirmed by qRT–PCR (right graph) (n = 3); data are presented as mean and s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001.
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Ohno, H., Shinoda, K., Ohyama, K. et al. EHMT1 controls brown adipose cell fate and thermogenesis through the PRDM16 complex. Nature 504, 163–167 (2013). https://doi.org/10.1038/nature12652
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