A PERK–miR-211 axis suppresses circadian regulators and protein synthesis to promote cancer cell survival

Abstract

The unfolded protein response (UPR) is a stress-activated signalling pathway that regulates cell proliferation, metabolism and survival. The circadian clock coordinates metabolism and signal transduction with light/dark cycles. We explore how UPR signalling interfaces with the circadian clock. UPR activation induces a 10 h phase shift in circadian oscillations through induction of miR-211, a PERK-inducible microRNA that transiently suppresses both Bmal1 and Clock, core circadian regulators. Molecular investigation reveals that miR-211 directly regulates Bmal1 and Clock via distinct mechanisms. Suppression of Bmal1 and Clock has the anticipated impact on expression of select circadian genes, but we also find that repression of Bmal1 is essential for UPR-dependent inhibition of protein synthesis and cell adaptation to stresses that disrupt endoplasmic reticulum homeostasis. Our data demonstrate that c-Myc-dependent activation of the UPR inhibits Bmal1 in Burkitt’s lymphoma, thereby suppressing both circadian oscillation and ongoing protein synthesis to facilitate tumour progression.

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Fig. 1: PERK-dependent regulation of circadian oscillation.
Fig. 2: Light/dark reversal triggers the UPR.
Fig. 3: miR-211 directly regulates Clock.
Fig. 4: miR-211 directly targets Bmal1 5′UTR.
Fig. 5: Antagonizing miR-211 restores circadian oscillation.
Fig. 6: Bmal1 is lost in Myc-driven tumours.
Fig. 7: Bmal1 loss is critical for cell survival following ER stress.
Fig. 8: Bmal1 loss is critical for PERK-dependent inhibition of protein synthesis.

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Acknowledgements

The authors thank J. Hogenesch (University of Cincinnati) for providing the U2OS BMAL1::Luc cell line. The authors thank Y. Koutalos (Medical University of South Carolina) for providing the animal darkroom facility, S. Mehrotra (Medical University of South Carolina) and C. Wallace Fugle (Medical University of South Carolina) for technique support, L.J. Valentijn (University of Amsterdam) for providing the SKNAS-NmycER cell line, and the Medical University of South Carolina pathology core research facility and Y. Shao for histological analysis. The authors also acknowledge assistance from the Division of Laboratory Animal Resource (DLAR) and the flow cytometry facility in the Medical University of South Carolina. This work was supported by National Institutes of Health grants P01CA165997 (to C.K., D.R., S.Y.F. and J.A.D.).

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J.A.D. developed the concept, designed experiments, analysed data and wrote the manuscript. Y.B. performed experiments, analysed data and prepared the manuscript. A.Y., N.C., B.J.A., F.T., A.T., A.O. and V.G. performed experiments. G.B.W. provided human samples of tonsils. K.E.A. carried out statistical analysis. S.M., C.V.D., D.R., C.K. and S.Y.F. provided intellectual input and reagents.

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Correspondence to J. Alan Diehl.

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Supplementary Table 1

Sequences for qPCR primers used in this study.

Supplementary Table 2

Information on antibodies used in this study.

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Bu, Y., Yoshida, A., Chitnis, N. et al. A PERK–miR-211 axis suppresses circadian regulators and protein synthesis to promote cancer cell survival. Nat Cell Biol 20, 104–115 (2018). https://doi.org/10.1038/s41556-017-0006-y

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