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The orphan nuclear receptor RORα regulates circadian transcription of the mammalian core-clock Bmal1

Nature Structural & Molecular Biology volume 12, pages 441–448 (2005)Cite this article

Abstract

The PAS (PER-ARNT-SIM) helix-loop-helix transcription factor BMAL1 (also known as MOP3) is an essential component of the circadian pacemaker in mammals. Here we show that the retinoic acid receptor–related orphan receptor RORα (NR1F1) directly activates transcription of Bmal1 through two conserved RORα response elements that are required for cell-autonomous transcriptional oscillation of Bmal1 mRNA. Positive involvement of RORα in generation of the Bmal1 circadian oscillation was verified by behavioral analyses of RORα-deficient staggerer mice that showed aberrant locomotor activity and unstable rhythmicity. In cultured cells, loss of endogenous RORα protein resulted in a dampened circadian rhythm of Bmal1 transcription, further indicating that RORα is a functional component of the cell-autonomous core circadian clock. These results indicate that RORα acts to promote Bmal1 transcription, thereby maintaining a robust circadian rhythm.

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Figure 1: Temporal mRNA expression patterns of Rev-erbα and Rorα, and effects of overexpression of REV-ERB or/and ROR on Bmal1 transcription.
Figure 2: Effect of a dominant-negative RORα on the circadian transcription of Bmal1 Transcriptional oscillation of Bmal1 was monitored by using a cell culture–based luminescent reporter assay, in the presence or absence of β-galactosidase or a dominant-negative form of RORα.
Figure 3: Region on the Bmal1 promoter required for RORα-mediated transactivation and transcriptional oscillation.
Figure 4: Temporal pattern of RORα binding to the Bmal1 promoter region in pull-down assays.
Figure 5: Circadian phenotypes of RORα-deficient staggerer mice.
Figure 6: Effect of RORα deficiency on the cell-autonomous circadian transcription of Bmal1.
Figure 7: Schematic model representing RORα-mediated control of cell-autonomous Bmal1 oscillations.

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Acknowledgements

We thank Y. Sakakida, S. Yoshiba, S. Tsuboi, C. Matsubara and T. Ichise for their expert technical assistance, and all the members of the laboratory for help, reagents and discussions. We express our great appreciation to J. Dunlap, A. Sehgal, G. Block and R. Evans for their reviewing the manuscript and also thank T. Mamine, E. Nishida and K. Tanaka for general support. We are grateful to T. Kono for the siRNA expression vector and S. Ishii for the pDECAP vector. This work was supported in part by research grants from the Japan Ministry of Education, Culture, Sports, Science and Technology and Sony Corporation. The support of fellowships from the Japan Society for the Promotion of Science (M.A.) is also acknowledged.

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  1. Osaka Bioscience Institute, 6-2-4 Furuedai, Suita, 565-0874, Osaka, Japan

    Makoto Akashi & Toru Takumi

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Correspondence to Toru Takumi.

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Supplementary information

Supplementary Fig. 1

Effect of RORα antisense on long dsRNA. (PDF 347 kb)

Supplementary Fig. 2

Effect of cobalt chloride on BMAL1 oscillation. (PDF 134 kb)

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Akashi, M., Takumi, T. The orphan nuclear receptor RORα regulates circadian transcription of the mammalian core-clock Bmal1. Nat Struct Mol Biol 12, 441–448 (2005). https://doi.org/10.1038/nsmb925

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