EHMT1 controls brown adipose cell fate and thermogenesis through the PRDM16 complex


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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Figure 1: Identification of EHMT1 in the PRDM16 transcriptional complex.
Figure 2: EHMT1 is required for BAT versus muscle lineage specification.
Figure 3: EHMT1 controls BAT thermogenesis through stabilizing PRDM16 protein.
Figure 4: EHMT1 deficiency in BAT causes obesity and insulin resistance.


  1. 1

    Seale, P. et al. PRDM16 controls a brown fat/skeletal muscle switch. Nature 454, 961–967 (2008)

    CAS  ADS  Article  Google Scholar 

  2. 2

    Kajimura, S. et al. Initiation of myoblast to brown fat switch by a PRDM16–C/EBP-β transcriptional complex. Nature 460, 1154–1158 (2009)

    CAS  ADS  Article  Google Scholar 

  3. 3

    Nedergaard, J., Bengtsson, T. & Cannon, B. Unexpected evidence for active brown adipose tissue in adult humans. Am. J. Physiol. 293, E444–E452 (2007)

    CAS  Google Scholar 

  4. 4

    van Marken Lichtenbelt, W. D. et al. Cold-activated brown adipose tissue in healthy men. N. Engl. J. Med. 360, 1500–1508 (2009)

    CAS  Article  Google Scholar 

  5. 5

    Saito, M. et al. High incidence of metabolically active brown adipose tissue in healthy adult humans: effects of cold exposure and adiposity. Diabetes 58, 1526–1531 (2009)

    CAS  Article  Google Scholar 

  6. 6

    Virtanen, K. A. et al. Functional brown adipose tissue in healthy adults. N. Engl. J. Med. 360, 1518–1525 (2009)

    CAS  Article  Google Scholar 

  7. 7

    Atit, R. et al. β-catenin activation is necessary and sufficient to specify the dorsal dermal fate in the mouse. Dev. Biol. 296, 164–176 (2006)

    CAS  Article  Google Scholar 

  8. 8

    Timmons, J. A. et al. Myogenic gene expression signature establishes that brown and white adipocytes originate from distinct cell lineages. Proc. Natl Acad. Sci. USA 104, 4401–4406 (2007)

    CAS  ADS  Article  Google Scholar 

  9. 9

    Kajimura, S., Seale, P. & Spiegelman, B. M. Transcriptional control of brown fat development. Cell Metab. 11, 257–262 (2010)

    CAS  Article  Google Scholar 

  10. 10

    Shing, D. C. et al. Overexpression of sPRDM16 coupled with loss of p53 induces myeloid leukemias in mice. J. Clin. Invest. 117, 3696–3707 (2007)

    CAS  PubMed  PubMed Central  Google Scholar 

  11. 11

    Pinheiro, I. et al. Prdm3 and Prdm16 are H3K9me1 methyltransferases required for mammalian heterochromatin integrity. Cell 150, 948–960 (2012)

    CAS  Article  Google Scholar 

  12. 12

    Tachibana, M. et al. Histone methyltransferases G9a and GLP form heteromeric complexes and are both crucial for methylation of euchromatin at H3–K9. Genes Dev. 19, 815–826 (2005)

    CAS  Article  Google Scholar 

  13. 13

    Kleefstra, T. et al. Disruption of the gene euchromatin histone methyl transferase1 (Eu-HMTase1) is associated with the 9q34 subtelomeric deletion syndrome. J. Med. Genet. 42, 299–306 (2005)

    CAS  Article  Google Scholar 

  14. 14

    Cormier-Daire, V. et al. Cryptic terminal deletion of chromosome 9q34: a novel cause of syndromic obesity in childhood? J. Med. Genet. 40, 300–303 (2003)

    CAS  Article  Google Scholar 

  15. 15

    Willemsen, M. H. et al. Update on Kleefstra syndrome. Mol. Syndromol. 2, 202–212 (2012)

    CAS  PubMed  PubMed Central  Google Scholar 

  16. 16

    Kajimura, S. et al. Regulation of the brown and white fat gene programs through a PRDM16/CtBP transcriptional complex. Genes Dev. 22, 1397–1409 (2008)

    CAS  Article  Google Scholar 

  17. 17

    Schaefer, A. et al. Control of cognition and adaptive behavior by the GLP/G9a epigenetic suppressor complex. Neuron 64, 678–691 (2009)

    CAS  Article  Google Scholar 

  18. 18

    Tachibana, M., Matsumura, Y., Fukuda, M., Kimura, H. & Shinkai, Y. G9a/GLP complexes independently mediate H3K9 and DNA methylation to silence transcription. EMBO J. 27, 2681–2690 (2008)

    CAS  Article  Google Scholar 

  19. 19

    Eguchi, J. et al. Transcriptional control of adipose lipid handling by IRF4. Cell Metab. 13, 249–259 (2011)

    CAS  Article  Google Scholar 

  20. 20

    Almind, K., Manieri, M., Sivitz, W. I., Cinti, S. & Kahn, C. R. Ectopic brown adipose tissue in muscle provides a mechanism for differences in risk of metabolic syndrome in mice. Proc. Natl Acad. Sci. USA 104, 2366–2371 (2007)

    CAS  ADS  Article  Google Scholar 

  21. 21

    Cannon, B. & Nedergaard, J. Nonshivering thermogenesis and its adequate measurement in metabolic studies. J. Exp. Biol. 214, 242–253 (2011)

    Article  Google Scholar 

  22. 22

    Ouellet, V. et al. Outdoor temperature, age, sex, body mass index, and diabetic status determine the prevalence, mass, and glucose-uptake activity of 18F-FDG-detected BAT in humans. J. Clin. Endocrinol. Metab. 96, 192–199 (2012)

    Article  Google Scholar 

  23. 23

    Wu, Q. et al. Fatty acid transport protein 1 is required for nonshivering thermogenesis in brown adipose tissue. Diabetes 55, 3229–3237 (2006)

    CAS  Article  Google Scholar 

  24. 24

    Feldmann, H. M., Golozoubova, V., Cannon, B. & Nedergaard, J. UCP1 ablation induces obesity and abolishes diet-induced thermogenesis in mice exempt from thermal stress by living at thermoneutrality. Cell Metab. 9, 203–209 (2009)

    CAS  Article  Google Scholar 

  25. 25

    Yoneshiro, T. et al. Impact of UCP1 and beta3AR gene polymorphisms on age-related changes in brown adipose tissue and adiposity in humans. Int. J. Obes. 37, 993–998 (2013)

    CAS  Article  Google Scholar 

  26. 26

    Huh, M. S., Parker, M. H., Scime, A., Parks, R. & Rudnicki, M. A. Rb is required for progression through myogenic differentiation but not maintenance of terminal differentiation. J. Cell Biol. 166, 865–876 (2004)

    CAS  Article  Google Scholar 

  27. 27

    Mao, X. et al. APPL1 binds to adiponectin receptors and mediates adiponectin signalling and function. Nature Cell Biol. 8, 516–523 (2006)

    CAS  Article  Google Scholar 

  28. 28

    Liisberg Aune, U., Ruiz, L. & Kajimura, S. Isolation and differentiation of stromal vascular cells to beige/brite cells. J Visual. Exp. e50191 (2013)

  29. 29

    Takahashi, A. et al. DNA damage signaling triggers degradation of histone methyltransferases through APC/C(Cdh1) in senescent cells. Mol. Cell 45, 123–131 (2012)

    CAS  Article  Google Scholar 

  30. 30

    Kurn, N. et al. Novel isothermal, linear nucleic acid amplification systems for highly multiplexed applications. Clin. Chem. 51, 1973–1981 (2005)

    CAS  Article  Google Scholar 

  31. 31

    Trapnell, C. et al. Differential analysis of gene regulation at transcript resolution with RNA-seq. Nature Biotechnol. 31, 46–53 (2013)

    CAS  Article  Google Scholar 

  32. 32

    Huang, W., Sherman, B. T. & Lempicki, R. A. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nature Protocols 4, 44–57 (2009)

    CAS  Article  Google Scholar 

  33. 33

    Kersey, P. J. et al. The International Protein Index: an integrated database for proteomics experiments. Proteomics 4, 1985–1988 (2004)

    CAS  Article  Google Scholar 

  34. 34

    Elias, J. E. & Gygi, S. P. Target-decoy search strategy for increased confidence in large-scale protein identifications by mass spectrometry. Nature Methods 4, 207–214 (2007)

    CAS  Article  Google Scholar 

  35. 35

    Seale, P. et al. Transcriptional control of brown fat determination by PRDM16. Cell Metab. 6, 38–54 (2007)

    CAS  Article  Google Scholar 

  36. 36

    Ohno, H., Shinoda, K., Spiegelman, B. M. & Kajimura, S. PPARγ agonists induce a white-to-brown fat conversion through stabilization of PRDM16 protein. Cell Metab. 15, 395–404 (2012)

    CAS  Article  Google Scholar 

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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.

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S.K. and H.O. conceived and designed the experiments. All authors performed the experiments and analysed the data. S.K. and H.O. wrote the paper.

Corresponding author

Correspondence to Shingo Kajimura.

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The authors declare no competing financial interests.

Extended data figures and tables

Extended Data Figure 1 EHMT1 regulates endogenous PRDM16 protein expression in vivo.

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.

Extended Data Figure 3 EHMT1 is required for beige/brite cell development.

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).

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