Abstract
The circadian clock regulates behavioural and physiological processes in a 24-h cycle. The nuclear receptors REV-ERBα and REV-ERBβ are involved in the cell-autonomous circadian transcriptional/translational feedback loops as transcriptional repressors. A number of studies have also demonstrated a pivotal role of REV-ERBs in regulation of metabolic, neuronal, and inflammatory functions including bile acid metabolism, lipid metabolism, and production of inflammatory cytokines. Given the multifunctional role of REV-ERBs, it is important to elucidate the mechanism through which REV-ERBs exert their functions. To this end, we established a Rev-erbα/Rev-erbβ double-knockout mouse embryonic stem (ES) cell model and analyzed the circadian clock and clock-controlled output gene expressions. A comprehensive mRNA-seq analysis revealed that the double knockout of both Rev-erbα and Rev-erbβ does not abrogate expression rhythms of E-box-regulated core clock genes but drastically changes a diverse set of other rhythmically-expressed output genes. Of note, REV-ERBα/β deficiency does not compromise circadian expression rhythms of PER2, while REV-ERB target genes, Bmal1 and Npas2, are significantly upregulated. This study highlight the relevance of REV-ERBs as pivotal output mediators of the mammalian circadian clock.
Highlights
The circadian clock regulates behavioural and physiological processes in a 24-h cycle
Translational frameshifts were confirmed by sequencing of cDNA (Supplementary Fig. S1) and mRNAs including the CRISPR-targeted exon were not detected by quantitative PCR (Fig. 1b), indicating successful establishment of Rev-erbα/Rev-erbβ double knockout mouse embryonic stem (mES) cells
To evaluate the impact of REV-ERBs deficiency on global gene expression, the cells were differentiated with the embryoid body (EB) formation method and temporal RNA sequencing (RNA-seq) analysis with RNA samples at 4-h intervals over 2 days were performed
Summary
The circadian clock regulates behavioural and physiological processes in a 24-h cycle. Given the multifunctional role of REV-ERBs, it is important to elucidate the mechanism through which REVERBs exert their functions To this end, we established a Rev-erbα/Rev-erbβ double-knockout mouse embryonic stem (ES) cell model and analyzed the circadian clock and clock-controlled output gene expressions. It has been reported that liver-specific deficiency and the induced deficiency of both Rev-erbα and Rev-erbβ in adult mice results in altered circadian gene expressions and disrupted behavioral rhythms, respectively[8]. These observations suggest the redundant and essential role of REV-ERBα and REV-ERBβ in circadian clock regulation. W mRNA expression level b sequence, respectively (see Methods). (b) Relative expression levels of Rev-erbα and Rev-erbβ mRNA including
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
