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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Diehl, J. A., Fuchs, S. Y. & Koumenis, C. The cell biology of the unfolded protein response. Gastroenterology 141, 38–41.e2 (2011).
Tirasophon, W., Welihinda, A. A. & Kaufman, R. J. A stress response pathway from the endoplasmic reticulum to the nucleus requires a novel bifunctional protein kinase/endoribonuclease (Ire1p) in mammalian cells. Genes Dev. 12, 1812–1824 (1998).
Yoshida, H., Haze, K., Yanagi, H., Yura, T. & Mori, K. Identification of the cis-acting endoplasmic reticulum stress response element responsible for transcriptional induction of mammalian glucose-regulated proteins. Involvement of basic leucine zipper transcription factors. J. Biol. Chem. 273, 33741–33749 (1998).
Lee, A. H., Iwakoshi, N. N. & Glimcher, L. H. XBP-1 regulates a subset of endoplasmic reticulum resident chaperone genes in the unfolded protein response. Mol. Cell Biol. 23, 7448–7459 (2003).
Calfon, M. et al. IRE1 couples endoplasmic reticulum load to secretory capacity by processing the XBP-1 mRNA. Nature 415, 92–96 (2002).
Yoshida, H., Matsui, T., Yamamoto, A., Okada, T. & Mori, K. XBP1 mRNA is induced by ATF6 and spliced by IRE1 in response to ER stress to produce a highly active transcription factor. Cell 107, 881–891 (2001).
Shi, Y. et al. Identification and characterization of pancreatic eukaryotic initiation factor 2 alpha-subunit kinase, PEK, involved in translational control. Mol. Cell Biol. 18, 7499–7509 (1998).
Harding, H. P., Zhang, Y. & Ron, D. Protein translation and folding are coupled by an endoplasmic-reticulum-resident kinase. Nature 397, 271–274 (1999).
Pytel, D., Majsterek, I. & Diehl, J. A. Tumor progression and the different faces of the PERK kinase. Oncogene 35, 1207–1215 (2016).
Bobrovnikova-Marjon, E. et al. PERK promotes cancer cell proliferation and tumor growth by limiting oxidative DNA damage. Oncogene 29, 3881–3895 (2010).
Hart, L. S. et al. ER stress-mediated autophagy promotes Myc-dependent transformation and tumor growth. J. Clin. Invest. 122, 4621–4634 (2012).
Pytel, D. et al. PERK is a haploinsufficient tumor suppressor: gene dose determines tumor-suppressive versus tumor promoting properties of PERK in melanoma. PLoS Genet. 12, e1006518 (2016).
Bhattacharya, S. et al. Anti-tumorigenic effects of type 1 interferon are subdued by integrated stress responses. Oncogene 32, 4214–4221 (2013).
Harding, H. P., Zhang, Y., Bertolotti, A., Zeng, H. & Ron, D. PERK is essential for translational regulation and cell survival during the unfolded protein response. Mol. Cell 5, 897–904 (2000).
Zhang, P. et al. The PERK eukaryotic initiation factor 2α kinase is required for the development of the skeletal system, postnatal growth, and the function and viability of the pancreas. Mol. Cell. Biol. 22, 3864–3874 (2002).
Gao, Y. et al. PERK is required in the adult pancreas and is essential for maintenance of glucose homeostasis. Mol. Cell. Biol. 32, 5129–5139 (2012).
Lipton, J. O. et al. The circadian protein BMAL1 regulates translation in response to S6K1-mediated phosphorylation. Cell 161, 1138–1151 (2015).
Bass, J. & Takahashi, J. S. Circadian integration of metabolism and energetics. Science 330, 1349–1354 (2010).
Bu, Y. & Diehl, J. A. PERK integrates oncogenic signaling and cell survival during cancer development. J. Cell. Physiol. 231, 2088–2096 (2016).
Baggs, J. E. et al. Network features of the mammalian circadian clock. PLoS Biol. 7, e52 (2009).
Axten, J. M. et al. Discovery of 7-methyl-5-(1-{[3-(trifluoromethyl)phenyl]acetyl}-2,3-dihydro-1H-indol-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (GSK2606414), a potent and selective first-in-class inhibitor of protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK). J. Med. Chem. 55, 7193–7207 (2012).
Chitnis, N. S. et al. miR-211 is a prosurvival microRNA that regulates chop expression in a PERK-dependent manner. Mol. Cell 48, 353–364 (2012).
Harfe, B. D., McManus, M. T., Mansfield, J. H., Hornstein, E. & Tabin, C. J. The RNaseIII enzyme Dicer is required for morphogenesis but not patterning of the vertebrate limb. Proc. Natl Acad. Sci. USA 102, 10898–10903 (2005).
Verdel, A. et al. RNAi-mediated targeting of heterochromatin by the RITS complex. Science 303, 672–676 (2004).
Orom, U. A. & Lund, A. H. Isolation of microRNA targets using biotinylated synthetic microRNAs. Methods 43, 162–165 (2007).
Barna, M. et al. Suppression of Myc oncogenic activity by ribosomal protein haploinsufficiency. Nature 456, 971–975 (2008).
Ruggero, D. The role of Myc-induced protein synthesis in cancer. Cancer Res. 69, 8839–8843 (2009).
Altman, B. J. et al. MYC disrupts the circadian clock and metabolism in cancer cells. Cell Metab. 22, 1009–1019 (2015).
Garten, A. et al. Physiological and pathophysiological roles of NAMPT and NAD metabolism. Nat. Rev. Endocrinol. 11, 535–546 (2015).
Atwood, A. et al. Cell-autonomous circadian clock of hepatocytes drives rhythms in transcription and polyamine synthesis. Proc. Natl Acad. Sci. USA 108, 18560–18565 (2011).
Eckel-Mahan, K. L. et al. Reprogramming of the circadian clock by nutritional challenge. Cell 155, 1464–1478 (2013).
Li, Y. et al. PRMT5 is required for lymphomagenesis triggered by multiple oncogenic drivers. Cancer Discov. 5, 288–303 (2015).
Basso, K. et al. Reverse engineering of regulatory networks in human B cells. Nat. Genet. 37, 382–390 (2005).
Avivar-Valderas, A. et al. Regulation of autophagy during ECM detachment is linked to a selective inhibition of mTORC1 by PERK. Oncogene 32, 4932–4940 (2013).
Gyorffy, B., Surowiak, P., Budczies, J. & Lanczky, A. Online survival analysis software to assess the prognostic value of biomarkers using transcriptomic data in non-small-cell lung cancer. PLoS ONE 8, e82241 (2013).
Delmore, J. E. et al. BET bromodomain inhibition as a therapeutic strategy to target c-Myc. Cell 146, 904–917 (2011).
Belinsky, M. G., Chen, Z. S., Shchaveleva, I., Zeng, H. & Kruh, G. D. Characterization of the drug resistance and transport properties of multidrug resistance protein 6 (MRP6, ABCC6). Cancer Res. 62, 6172–6177 (2002).
Valentijn, L. J. et al. Inhibition of a new differentiation pathway in neuroblastoma by copy number defects of N-myc, Cdc42, and nm23 genes. Cancer Res. 65, 3136–3145 (2005).
Ushmorov, A. et al. N-myc augments death and attenuates protective effects of Bcl-2 in trophically stressed neuroblastoma cells. Oncogene 27, 3424–3434 (2008).
Yoshida, A., Lee, E. K. & Diehl, J. A. Induction of therapeutic senescence in vemurafenib-resistant melanoma by extended inhibition of CDK4/6. Cancer Res. 76, 2990–3002 (2016).
Xu, Z. et al. miR-216b regulation of c-Jun mediates GADD153/CHOP-dependent apoptosis. Nat. Commun. 7, 11422 (2016).
Aggarwal, P. et al. Nuclear cyclin D1/CDK4 kinase regulates CUL4 expression and triggers neoplastic growth via activation of the PRMT5 methyltransferase. Cancer Cell 18, 329–340 (2010).
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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Supplementary Information
Supplementary Figures 1–8, plus legends and references.
Supplementary Table 1
Sequences for qPCR primers used in this study.
Supplementary Table 2
Information on antibodies used in this study.
Supplementary Table 3
Statistics Source Data.
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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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DOI: https://doi.org/10.1038/s41556-017-0006-y
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