Regulation of circadian behaviour and metabolism by REV-ERB-α and REV-ERB-β - PubMed (original) (raw)

. 2012 Mar 29;485(7396):123-7.

doi: 10.1038/nature11048.

Xuan Zhao, Megumi Hatori, Ruth T Yu, Grant D Barish, Michael T Lam, Ling-Wa Chong, Luciano DiTacchio, Annette R Atkins, Christopher K Glass, Christopher Liddle, Johan Auwerx, Michael Downes, Satchidananda Panda, Ronald M Evans

Affiliations

Regulation of circadian behaviour and metabolism by REV-ERB-α and REV-ERB-β

Han Cho et al. Nature. 2012.

Abstract

The circadian clock acts at the genomic level to coordinate internal behavioural and physiological rhythms via the CLOCK-BMAL1 transcriptional heterodimer. Although the nuclear receptors REV-ERB-α and REV-ERB-β have been proposed to form an accessory feedback loop that contributes to clock function, their precise roles and importance remain unresolved. To establish their regulatory potential, we determined the genome-wide cis-acting targets (cistromes) of both REV-ERB isoforms in murine liver, which revealed shared recognition at over 50% of their total DNA binding sites and extensive overlap with the master circadian regulator BMAL1. Although REV-ERB-α has been shown to regulate Bmal1 expression directly, our cistromic analysis reveals a more profound connection between BMAL1 and the REV-ERB-α and REV-ERB-β genomic regulatory circuits than was previously suspected. Genes within the intersection of the BMAL1, REV-ERB-α and REV-ERB-β cistromes are highly enriched for both clock and metabolic functions. As predicted by the cistromic analysis, dual depletion of Rev-erb-α and Rev-erb-β function by creating double-knockout mice profoundly disrupted circadian expression of core circadian clock and lipid homeostatic gene networks. As a result, double-knockout mice show markedly altered circadian wheel-running behaviour and deregulated lipid metabolism. These data now unite REV-ERB-α and REV-ERB-β with PER, CRY and other components of the principal feedback loop that drives circadian expression and indicate a more integral mechanism for the coordination of circadian rhythm and metabolism.

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Figures

Figure 1

Figure 1. Cistromic analyses of REV-ERBα and REV-ERBβ in liver

a, De novo HOMER motif analysis of in vivo REV-ERBα and REV-ERBβ binding. b, Venn diagram depicting the unique and common REV- ERBα and REV-ERBβ bound peaks. c, Commonly bound REV-ERBα and REV-ERBβ peaks are enriched for genes involved in lipid metabolism and associated with PPARs. d, REV-ERBα, REV-ERBβ and BMAL1binding at canonical circadian clock genes. Left axis indicates tag counts. e, BMAL1 cistrome significantly overlaps with REV-ERBα and REV-ERBβ. Examples of Clock related genes in overlap are listed and selected peaks shown in Supplementary Figure 4.

Figure 2

Figure 2. Circadian gene expression of many canonical core clock genes and output genes are disrupted in Livers of Rev-erbαlox/lox Rev-erbβlox/lox Albumin-Cre (L-DKO) mice

The expression levels of a, Rev-erbα, b, Rev-erbβ, c-f, canonical core clock genes (Cry1, Clock, Bmal1 and Per2) g-i, presumed output genes (PoR, PPARα and Sco2) in livers from L-DKO (Albumin-Cre positive, red labels) and wildtype (Albumin-Cre negative, black labels) mice. Livers (n=3) were harvested at each indicated ZT under 12-hour light:dark cycle. QPCR was performed in technical triplicates. Relative Unit (RU) normalized with 36B4. Error bars indicate standard error of the mean, statistical significance determined by Student t-test (* p<0.05, ** p<0.01, *** p<0.001).

Figure 3

Figure 3. Broad disruption of circadian transcriptome in the absence of Rev-erbα and Rev-erbβ

a, Heatmap of genes with circadian expression in wild-type (left panel) and L-DKO (right panel) livers. 1227 unique accession numbers were selected based on fdr < 0.05. b, Genes expressed in a circadian manner that lose rhythm are highly associated with circadian and energy homeostasis functions as assessed by KEGG Pathway analysis.

Figure 4

Figure 4. Loss of both Rev-erbα and Rev-erbβ results in disrupted circadian wheel-running behavior and metabolic shift

a-c, Voluntary locomotor activity of wildtype, Rev-erbα−/−, Rev-erbβ−/−, and Rev-erbα−/−Rev-erbβ−/− (iDKO) mice. a, Actograms showing wheel-running activity in constant darkness after prior entrainment in light/dark. b, Activity profiles during light dark cycles. c, Chi-square periodogram of the initial 20 days in constant darkness. (n=5–9 for each mutant strain, n= 5–6 littermate controls). Representative actograms from individual mice are shown. d, Triglyceride (n=6), fasting glucose (n=6) and free fatty acid (n=6) levels in iDKO and wildtype mice. e, Respiratory exchange ratio (RER) for wildtype (black) and iDKO (red) mice (n=4). f, Model depicting the activating (Clock/BMAL1) and repressive (REV-ERBα/REV-ERBβ) transcriptional complexes whose coordinate actions generate rhythmic gene expression.

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