Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2018 Apr 19;9(1):1566.
doi: 10.1038/s41467-018-03868-8.

Histone demethylase JMJD1A coordinates acute and chronic adaptation to cold stress via thermogenic phospho-switch

Yohei Abe  1 , Yosuke Fujiwara  1 , Hiroki Takahashi  1 , Yoshihiro Matsumura  1 , Tomonobu Sawada  1 , Shuying Jiang  2 , Ryo Nakaki  3   4 , Aoi Uchida  1 , Noriko Nagao  1 , Makoto Naito  5 , Shingo Kajimura  6 , Hiroshi Kimura  7 , Timothy F Osborne  8 , Hiroyuki Aburatani  4 , Tatsuhiko Kodama  9 , Takeshi Inagaki  10   11 , Juro Sakai  12   13
Affiliations

Histone demethylase JMJD1A coordinates acute and chronic adaptation to cold stress via thermogenic phospho-switch

Yohei Abe et al. Nat Commun. .

Abstract

In acute cold stress in mammals, JMJD1A, a histone H3 lysine 9 (H3K9) demethylase, upregulates thermogenic gene expressions through β-adrenergic signaling in brown adipose tissue (BAT). Aside BAT-driven thermogenesis, mammals have another mechanism to cope with long-term cold stress by inducing the browning of the subcutaneous white adipose tissue (scWAT). Here, we show that this occurs through a two-step process that requires both β-adrenergic-dependent phosphorylation of S265 and demethylation of H3K9me2 by JMJD1A. The histone demethylation-independent acute Ucp1 induction in BAT and demethylation-dependent chronic Ucp1 expression in beige scWAT provides complementary molecular mechanisms to ensure an ordered transition between acute and chronic adaptation to cold stress. JMJD1A mediates two major signaling pathways, namely, β-adrenergic receptor and peroxisome proliferator-activated receptor-γ (PPARγ) activation, via PRDM16-PPARγ-P-JMJD1A complex for beige adipogenesis. S265 phosphorylation of JMJD1A, and the following demethylation of H3K9me2 might prove to be a novel molecular target for the treatment of metabolic disorders, via promoting beige adipogenesis.

PubMed Disclaimer

Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
Ucp1 expression and H3K9me2 levels in adipose tissues and BAT thermogenic function in Jmjd1a-S265AKI/KI mice . a Ucp1 mRNA (n = 3) (left) and protein levels detected by immunoblotting (IB) (right) in BAT, and scWAT of mice exposed to 4 °C for 6 h. b H3K9me2 immunoblotting using purified histones from adipose tissues of mice housed at RT. c H3K9me2 ChIP-qPCR in BAT, and scWAT of mice exposed to 4 °C for 6 h (n = 3). d NE-induced Ucp1 mRNA levels in im-BATs stably expressing WT or indicated mutants of hJMJD1A. The relative quantity in im-BATs expressing WT-JMJD1A on day 8 before NE treatment (0 h) is defined as 1. e Ucp1 mRNA expressions (n = 3) (left) and proteins (right) in BAT, and scWAT from mice exposed to 30 °C or 4 °C for 1 week. Uncropped images of the blots (a, b, e) are shown in Supplementary Fig. 8. f, g H3K9me2 ChIP-qPCR in scWAT from mice placed at 4 °C for 1 week (n = 4) (f) and in im-scWATs during beige adipogenesis (mean ± s.e.m. of three technical replicates) (g). h Bi-phasic Ucp1 expressions during cold exposure in BAT and scWAT. Mice were exposed to 4 °C for the indicated time, and Ucp1 mRNAs in BAT and scWATs were quantified by qPCR (n = 3). The data were converted to copy number per ng of total RNA. i In BAT, Ucp1 locus is in euchromatin (left), while in scWAT, it is in heterochromatin with H3K9me2 (right). In BAT, cold exposure leads to acute induction of Ucp1 mRNA through the mechanisms independent of H3K9me2 demethylation (left). In scWAT, H3K9me2 at Ucp1 gene locus needs to be removed for beige adipogenesis (right bottom). j Cold intolerance in Jmjd1a-S265AKI/KI mice (n = 6 per genotype group). Shown is the body temperature of 9-week-old mice at different times after cold exposure (4 °C). k Impaired NE-induced activation of selective genes in BAT (n = 6 per genotype group). l, m Reduced NE-induced mitochondrial respiration (l) and NE-induced glycerol release (m) in primary brown adipocytes from Jmjd1a-S265AKI/KI mice. Data are mean ± s.e.m. of five technical replicates (l) and three independent experiments (m). Data are mean ± s.e.m. a, c, e, f, j, k Student’s t test (a, e, f, j, l, m) or analysis of variance, followed by Tukey’s post hoc comparison (k) were performed for comparisons. *P < 0.05, **P < 0.01, and ***P < 0.005
Fig. 2
Fig. 2
Phospho-S265 JMJD1A induces beige biogenesis. a qPCR analysis demonstrates decreased expression of beige-selective genes in scWAT from Jmjd1a-S265AKI/KI mice exposed to 30 °C or 4 °C for 1 week (n = 5 per genotype group). b Immunoblot analysis of UCP1 and PPARγ in tissue homogenates of scWAT from mice presented in a. Uncropped images of the blots are shown in Supplementary Fig. 8. c Haematoxylin and eosin (H&E) and UCP1 staining sections of scWAT from WT and Jmjd1a-S265AKI/KI mice exposed to chronic cold exposure (4 °C for 1 week) (scale bar, 100 μm). d NE-induced oxygen consumption rate (OCR) in mice exposed to chronic cold exposure (4 °C for 4 weeks) (n = 7 per genotype group) (left). OCR before and 30 min after NE treatment are analyzed (right) (n = 7). e OCR of scWAT from mice exposed to 30 °C or 4 °C for 1 week (WT: n = 3; Jmjd1a-S265AKI/KI: n = 4). Data are mean ± s.e.m. a, d, e Analysis of variance were performed followed by Tukey’s post hoc comparison in a. Student’s t test was performed for comparisons in d, e. *P < 0.05, **P < 0.01, and ***P < 0.005 were considered statistically significant
Fig. 3
Fig. 3
P-JMJD1A cell autonomously induces beige adipogenesis. a pSer265-JMJD1A protein levels in WT (+/+) and S265A knock-in whole-cell lysates (WCL) from scWAT cultures treated with NE or vehicle for 1 h. b Decreased beige-selective gene expressions in S265A knock-in scWAT cultures treated with NE (10 μM) for 2 h (mean ± s.e.m. of three technical replicates). ORO staining of indicated genotype of scWAT cultures (inset). c Immunoblot analysis using anti-UCP1, anti-PGC1α, anti-PRDM16, anti-PPARγ, or anti-total OXPHOS antibodies cocktail, using WCL from WT and S265A knock-in scWAT cultures. d MitoTracker staining in indicated genotype scWAT cultures (scale bar, 100 μm). e Mitochondrial DNA (mt-DNA) contents measured by qPCR in indicated scWAT cultures (mean ± s.e.m. of three independent experiments). f Electron micrographs of indicated genotype of scWAT cultures (bar, 1 μm). Mitochondria (M) and lipid droplets (L) are indicated. g The OCR of indicated scWAT cultures (left). The arrows indicate the time of addition for oligomycin (Oligo), FCCP, and rotenone/antimycin A (Rot/Anti). Basal, maximum, and uncoupled respiration were calculated (mean ± s.e.m. of five technical replicates) (right). h Glycerol release from indicated scWAT cultures after the treatment with NE for 3 h (mean ± s.e.m. of three independent experiments). i Increased expressions of beige-selective genes in S265D-hJMJD1A-transduced im-scWATs (mean ± s.e.m. of three technical replicates). ORO staining and MitoTracker staining (inset) (scale bar, 50 μm). j Immunoblotting with anti-UCP1, anti-PGC1α, anti-PPARγ, or anti-total OXPHOS antibodies cocktail using WCL from indicated im-scWATs. Uncropped images of the blots (a, c, j) are shown in Supplementary Fig. 8. k Mitochondrial DNA content measured by qPCR in indicated im-scWATs (mean ± s.e.m. of three technical replicates). Student’s t test was performed for comparisons in b, g, h. *P < 0.05, **P < 0.01, and ***P < 0.005 were considered statistically significant
Fig. 4
Fig. 4
β-Adrenergic signal is required for the induction of beige-selective genes mediated by PPARγ ligand. a WT-hJMJD1A-transduced or S265A-hJMJD1A-transduced im-scWATs were differentiated for beige adipogenesis in the presence or absence of propranolol (Pro), as schematically illustrated (top), and ORO staining was performed (bottom). b Whole-cell lysates (WCL) from WT-hJMJD1A-transduced or S265A-hJMJD1A-transduced im-scWATs differentiated under Pro (100 nM) plus or minus condition were subjected to immunoprecipitation (IP) with anti-V5 antibody, followed by immunoblot (IB) analysis with anti-P-JMJD1A (pSer265) antibody. c qPCR analysis of beige-selective genes and general adipogenic genes in WT-hJMJD1A-transduced or S265A-hJMJD1A-transduced im-scWATs under Pro plus or minus condition (mean ± s.e.m. of three technical replicates). d Immunoblotting with anti-UCP1 or anti-total OXPHOS antibodies cocktail using WCL from indicated viral transduced im-scWATs, differentiated under Pro plus or minus condition. e OCRs (basal, maximum, and uncoupled) of WT-hJMJD1A-transduced or S265A-hJMJD1A-transduced im-scWATs, differentiated under Pro plus or minus condition. Data are mean ± s.e.m. of five technical replicates. f, g Immunoblotting with anti-PRDM16, anti-P-JMJD1A, anti-V5, anti-PPARγ, or anti-PGC1α antibody following immunoprecipitation with anti-PRDM16 antibody (f) or with anti-V5 antibody for JMJD1A (g), from WCL of differentiated WT-hJMJD1A-transduced or S265A-hJMJD1A-transduced im-scWATs. Uncropped images of the blots (b, d, f, g) are shown in Supplementary Fig. 8. Analysis of variance was performed, followed by Tukey’s post hoc comparison in e. *P < 0.05, **P < 0.01, and ***P < 0.005 were considered statistically significant. h Schematic drawing of p265-JMJD1A-PPARγ-PGC1α-PRDM16 protein complex. Integration of β-adrenergic-cAMP signaling and the PPARγ ligand binding is mediated by JMJD1A through a mechanism where pS265-JMJD1A forms a complex with PGC1α, PRDM16, and PPARγ to mediate expressions of beige-selective genes
Fig. 5
Fig. 5
JMJD1A demethylates H3K9me2 on beige-selective genes. a RNA-seq heat map depicting expression ratio comparison between beige and white adipocytes, differentiated from im-scWATs under rosiglitazone (ROS) plus or minus condition (left). RNA-seq heat map of 411 beige-selective genes from the left panel depicts comparison of expression ratio between WT-hJMJD1A-transduced and S265A-hJMJD1A-transduced im-scWATs (right). Changes are log2 expression ratios of FPKM, as indicated in a color intensity scale. b, c qPCR analysis of beige-selective genes and Pparg in im-scWATs differentiated with or without ROS (b) or differentiated WT-hJMJD1A-transduced or S265A-hJMJD1A-transduced im-scWATs (c). d H3K9me2 ChIP-qPCR on beige-selective genes using scWAT from age-matched WT (+/+) and Jmjd1a-null (−/−) mice placed at 4 °C for 1 week (n = 4 per genotype group). e JMJD1A ChIP-qPCR on beige-selective genes during ROS-induced beige adipogenesis in im-scWATs. f ChIP-seq profiles for JMJD1A and PPARγ on Ucp1, Cidea, and Ppara genomic regions in differentiated im-scWATs (beige cells) and im-BATs (BAT cells). g, h ChIP-qPCR showing isoproterenol (ISO) treatment increased JMJD1A recruitment in differentiated im-scWATs (g) and the decrease of H3K9me2 levels in beige-selective genes in differentiated beige adipocytes by ROS was blunted by Pro treatment (h). i JMJD1A ChIP-qPCR on beige-selective genes in scWAT of WT and Jmjd1a-S265AKI/KI mice following 1-week cold exposure (WT: n = 3; Jmjd1a-S265AKI/KI: n = 6). j ChIP-qPCR showing the decrease of H3K9me2 levels on indicated beige-selective genes during beige adipogenesis is impaired in S265A-hJMJD1A-transduced im-scWATs. The signal in day 0 of differentiation is set as 1. Data are mean ± s.e.m. of three technical replicates in a representative experiments performed at least three times (b, c, e, g, h, j). Student’s t test was performed for comparisons in d, i. *P < 0.05 and **P < 0.01 were considered statistically significant
Fig. 6
Fig. 6
Demethylation activity is pivotal for beige-selective gene inductions. a Schematic representation of the domain structure of human JMJD1A. Phosphorylation site at S265 and Fe(II) binding site at H1120 are shown. b OCR in im-scWATs stably expressing WT-hJMJD1A, S265D-hJMJD1A, or S265D-H1120Y-hJMJD1A treated sequentially with oligomycin (Oligo), FCCP, and rotenone/antimycin A (Rot/Anti) (left). Basal, maximum, and uncoupled respiration calculated from the left (right). Data are mean ± s.e.m. of three technical replicates in a representative experiment. Analysis of variance were performed, followed by Tukey’s post hoc comparison. *P < 0.05, **P < 0.01, and ***P < 0.005 were considered statistically significant. c Heat map of mRNA levels determined by RNA-seq analysis of the indicated WT or mutant versions of hJMJD1A expressing im-scWATs. Changes are log2 expression (FPKM) ratios. For reference, a color intensity scale is included. Thirty-four genes that are beige-selective, S265A downregulated, S265D upregulated, and H1120Y downregulated (H3K9 demethylation-dependent) are listed (right). d qPCR analysis confirming RNA-seq shown in c. Mean ± s.e.m. of three technical replicates. e Hypothetical model. Cold stimulated β-adrenergic signal leads to phosphorylation of JMJD1A (Step 1), which triggers the formation of a PRDM16-PGC1α-PPARγ transcription complex that targets beige-selective genes (Step 1, top). JMJD1A then demethylates H3K9me2 to turn on the transcription of these genes in scWAT (Step 2, bottom)
Fig. 7
Fig. 7
Insulin resistance phenotype of Jmjd1a-S265AKI/KI mice. a Body weight changes. WT (+/+) and Jmjd1a-S265AKI/KI mice (WT: n = 6; Jmjd1a-S265AKI/KI: n = 8) were fed on HFD for 4 weeks at 30 °C, and then switched to 4 °C for 4 weeks. b, c Glucose tolerance test (GTT) (b) and insulin tolerance test (ITT) (c) in each genotype group mice fed on HFD after cold acclimation in a (WT: n = 6; Jmjd1a-S265AKI/KI: n = 7). d Assessment of insulin signaling as quantified by the phosphorylation of AKT-S473 in scWAT, BAT, or soleus from each genotype mice fed on HFD after cold acclimation presented in a. Uncropped images of the blots are shown in Supplementary Fig. 8. Data are mean ± s.e.m. (a–c) Student’s t test was performed for comparisons in a–c. *P < 0.05 and **P < 0.01 were considered statistically significant
Fig. 8
Fig. 8
Complementary mechanisms for thermogenic gene induction in acute and chronic cold stress via overlapping, but distinct mechanisms of JMJD1A. Brown fat cells mediate acute and robust thermogenic activation of Ucp1, while scWAT-derived beige fat cells contribute to an adaptive response against chronic cold exposure (top). The acute response in BAT requires a BAR-dependent phosphorylation of JMJD1A that facilitates long-range enhancer-promoter interactions and stimulate thermogenic gene expressions, but this does not require the intrinsic H3K9me2 demethylation activity of JMJD1A (left bottom). The chronic adaptation in beigeing requires both phosphorylation-dependent chromatin recruitment and H3K9me2 demethylation activity of JMJD1A (right bottom). These histone demethylation-independent acute Ucp1 induction in BAT and demethylation-dependent chronic Ucp1 expression in beige scWAT ensure an ordered transition between acute and chronic adaptation to cold stress. TXN transcription
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Wu J, et al. Beige adipocytes are a distinct type of thermogenic fat cell in mouse and human. Cell. 2012;150:366–376. doi: 10.1016/j.cell.2012.05.016. - DOI - PMC - PubMed
    1. Harms M, Seale P. Brown and beige fat: development, function and therapeutic potential. Nat. Med. 2013;19:1252–1263. doi: 10.1038/nm.3361. - DOI - PubMed
    1. Inagaki T, Sakai J, Kajimura S. Transcriptional and epigenetic control of brown and beige adipose cell fate and function. Nat. Rev. Mol. Cell. Biol. 2016;17:480–495. doi: 10.1038/nrm.2016.62. - DOI - PMC - PubMed
    1. Feldmann HM, 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. 2009;9:203–209. doi: 10.1016/j.cmet.2008.12.014. - DOI - PubMed
    1. Cannon B, Nedergaard J. Metabolic consequences of the presence or absence of the thermogenic capacity of brown adipose tissue in mice (and probably in humans) Int. J. Obes. (Lond.). 2010;34:S7–S16. doi: 10.1038/ijo.2010.177. - DOI - PubMed

Publication types

MeSH terms