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Ultradian hormone stimulation induces glucocorticoid receptor-mediated pulses of gene transcription

Nature Cell Biology volume 11, pages 1093–1102 (2009)Cite this article

Abstract

Studies on glucocorticoid receptor (GR) action typically assess gene responses by long-term stimulation with synthetic hormones. As corticosteroids are released from adrenal glands in a circadian and high-frequency (ultradian) mode, such treatments may not provide an accurate assessment of physiological hormone action. Here we demonstrate that ultradian hormone stimulation induces cyclic GR-mediated transcriptional regulation, or gene pulsing, both in cultured cells and in animal models. Equilibrium receptor-occupancy of regulatory elements precisely tracks the ligand pulses. Nascent RNA transcripts from GR-regulated genes are released in distinct quanta, demonstrating a profound difference between the transcriptional programs induced by ultradian and constant stimulation. Gene pulsing is driven by rapid GR exchange with response elements and by GR recycling through the chaperone machinery, which promotes GR activation and reactivation in response to the ultradian hormone release, thus coupling promoter activity to the naturally occurring fluctuations in hormone levels. The GR signalling pathway has been optimized for a prompt and timely response to fluctuations in hormone levels, indicating that biologically accurate regulation of gene targets by GR requires an ultradian mode of hormone stimulation.

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Figure 1: GR–MMTV promoter interactions in response to ultradian corticosterone fluctuations.
Figure 2: GR-chaperone cycle.
Figure 3: GFP–GR loading and dynamics at the MMTV promoter array during ultradian stimulation with natural and synthetic glucocorticoids.
Figure 4: Equilibrium association of GR with endogenous GR response elements (GREs).
Figure 5: Cyclic RNA Polymerase II loading and dynamics in response to ultradian corticosterone treatment.
Figure 6: Effects of ultradian corticosterone treatment on transcription of GR-regulated genes in 3134 cells.
Figure 7: Differential transcription outputs of ultradian and constant hormone treatments.
Figure 8: Pulsatile corticosterone treatment of adrenalectomized (ADX) rats and a model for gene pulsing.

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References

  1. Lightman, S. L. et al. Hypothalamic-pituitary-adrenal function. Arch. Physiol Biochem. 110, 90–93 (2002).

    Article  CAS  Google Scholar 

  2. Lightman, S. L. Patterns of exposure to glucocorticoid receptor ligand. Biochem. Soc. Trans. 34, 1117–1118 (2006).

    Article  CAS  Google Scholar 

  3. Droste, S. K. et al. Corticosterone levels in the brain show a distinct ultradian rhythm but a delayed response to forced swim stress. Endocrinology 149, 3244–3253 (2008).

    Article  CAS  Google Scholar 

  4. Young, E. A., Abelson, J. & Lightman, S. L. Cortisol pulsatility and its role in stress regulation and health. Front Neuroendocrinol. 25, 69–76 (2004).

    Article  CAS  Google Scholar 

  5. Lewis, J. G., Bagley, C. J., Elder, P. A., Bachmann, A. W. & Torpy, D. J. Plasma free cortisol fraction reflects levels of functioning corticosteroid-binding globulin. Clin. Chim. Acta 359, 189–194 (2005).

    Article  CAS  Google Scholar 

  6. McNally, J. G., Mueller, W. G., Walker, D., Wolford, R. G. & Hager, G. L. The glucocorticoid receptor: Rapid exchange with regulatory sites in living cells. Science 287, 1262–1265 (2000).

    Article  CAS  Google Scholar 

  7. Kramer, P. et al. Transcriptional state of the mouse mammary tumor virus promoter can effect topological domain size in vivo. J. Biol. Chem. 274, 28590–28597 (1999).

    Article  CAS  Google Scholar 

  8. Walker, D., Htun, H. & Hager, G. L. Using inducible vectors to study intracellular trafficking of GFP-tagged steroid/nuclear receptors in living cells. Methods 19, 386–393 (1999).

    Article  CAS  Google Scholar 

  9. Becker, M. et al. Dynamic behavior of transcription factors on a natural promoter in living cells. EMBO Rep. 3, 1188–1194 (2002).

    Article  CAS  Google Scholar 

  10. Stavreva, D. A., Muller, W. G., Hager, G. L., Smith, C. L. & McNally, J. G. Rapid glucocorticoid receptor exchange at a promoter is coupled to transcription and regulated by chaperones and proteasomes. Mol. Cell. Biol. 24, 2682–2697 (2004).

    Article  CAS  Google Scholar 

  11. Liu, J. & DeFranco, D. B. Chromatin recycling of glucocorticoid receptors: implications for multiple roles of heat shock protein 90. Mol. Endocrinol. 13, 355–365 (1999).

    Article  CAS  Google Scholar 

  12. Qi, M., Stasenko, L. J. & DeFranco, D. B. Recycling and desensitization of glucocorticoid receptors in v- mos transformed cells depend on the ability of nuclear receptors to modulate gene expression. Mol. Endocrinol. 4, 455–464 (1990).

    Article  CAS  Google Scholar 

  13. Munck, A. & Foley, R. Kinetics of glucocorticoid-receptor complexes in rat thymus cells. J. Steroid Biochem. 7, 1117–1122 (1976).

    Article  CAS  Google Scholar 

  14. Pratt, W. B., Galigniana, M. D., Harrell, J. M. & DeFranco, D. B. Role of hsp90 and the hsp90-binding immunophilins in signalling protein movement. Cell Signal. 16, 857–872 (2004).

    Article  CAS  Google Scholar 

  15. Freeman, B. C. & Yamamoto, K. R. Disassembly of transcriptional regulatory complexes by molecular chaperones. Science 296, 2232–2235 (2002).

    Article  CAS  Google Scholar 

  16. Freeman, B. C. & Yamamoto, K. R. Continuous recycling: a mechanism for modulatory signal transduction. Trends Biochem. Sci. 26, 285–290 (2001).

    Article  CAS  Google Scholar 

  17. Rayasam, G. V. et al. Ligand specific dynamics of the progesterone receptor in living cells and during chromatin remodeling in vitro. Mol. Cell. Biol. 25, 2406–2418 (2005).

    Article  CAS  Google Scholar 

  18. Klokk, T. I. et al. Ligand-specific dynamics of the androgen receptor at its response element in living cells. Mol. Cell Biol. 27, 1823–1843 (2007).

    Article  CAS  Google Scholar 

  19. John, S. et al. Interaction of the glucocorticoid receptor with the global chromatin landscape. Mol. Cell 29, 611–624 (2008).

    Article  CAS  Google Scholar 

  20. Sugaya, K., Vigneron, M. & Cook, P. R. Mammalian cell lines expressing functional RNA polymerase II tagged with the green fluorescent protein. J. Cell Sci. 113 (Pt 15), 2679–2683 (2000).

    CAS  PubMed  Google Scholar 

  21. Conway-Campbell, B. L. et al. Proteasome-dependent down-regulation of activated nuclear hippocampal glucocorticoid receptors determines dynamic responses to corticosterone. Endocrinology 148, 5470–5477 (2007).

    Article  CAS  Google Scholar 

  22. Metivier, R. et al. Estrogen receptor-α directs ordered, cyclical, and combinatorial recruitment of cofactors on a natural target promoter. Cell 115, 751–763 (2003).

    Article  CAS  Google Scholar 

  23. Kang, Z., Pirskanen, A., Janne, O. A. & Palvimo, J. J. Involvement of proteasome in the dynamic assembly of the androgen receptor transcription complex. J. Biol. Chem. 277, 48366–48371 (2002).

    Article  CAS  Google Scholar 

  24. Rothman, M. S. & Wierman, M. E. The role of gonadotropin releasing hormone in normal and pathologic endocrine processes. Curr. Opin. Endocrinol. Diabetes Obes. 14, 306–310 (2007).

    Article  CAS  Google Scholar 

  25. Thompson, N. M., Davies, J. S., Mode, A., Houston, P. A. & Wells, T. Pattern-dependent suppression of growth hormone (GH) pulsatility by ghrelin and GH-releasing peptide-6 in moderately GH-deficient rats. Endocrinology 144, 4859–4867 (2003).

    Article  CAS  Google Scholar 

  26. Chadwick, D. & Goode, J. A. Mechanisms and Biological Significance of Pulsatile Hormone Secretion (John Wiley & Sons Ltd., 2000).

    Book  Google Scholar 

  27. Belchetz, P. E., Plant, T. M., Nakai, Y., Keogh, E. J. & Knobil, E. Hypophysial responses to continuous and intermittent delivery of hypopthalamic gonadotropin-releasing hormone. Science 202, 631–633 (1978).

    Article  CAS  Google Scholar 

  28. Gore, A. C. GnRH: The master molecule of reproduction (Kluwer Academic Publishers, 2002).

    Book  Google Scholar 

  29. Geva-Zatorsky, N. et al. Oscillations and variability in the p53 system. Mol. Syst. Biol. 2, 2006 (2006).

    Article  Google Scholar 

  30. Batchelor, E., Mock, C. S., Bhan, I., Loewer, A. & Lahav, G. Recurrent initiation: a mechanism for triggering p53 pulses in response to DNA damage. Mol. Cell 30, 277–289 (2008).

    Article  CAS  Google Scholar 

  31. Nelson, D. E. et al. Oscillations in NF-kappaB signaling control the dynamics of gene expression. Science 306, 704–708 (2004).

    Article  CAS  Google Scholar 

  32. Karpova, T. S. et al. Concurrent fast and slow cycling of a transcriptional activator at an endogenous promoter. Science 319, 466–469 (2008).

    Article  CAS  Google Scholar 

  33. Dolmetsch, R. E., Xu, K. & Lewis, R. S. Calcium oscillations increase the efficiency and specificity of gene expression. Nature 392, 933–936 (1998).

    Article  CAS  Google Scholar 

  34. Cai, L., Dalal, C. K. & Elowitz, M. B. Frequency-modulated nuclear localization bursts coordinate gene regulation. Nature 455, 485–490 (2008).

    Article  CAS  Google Scholar 

  35. Chubb, J. R., Trcek, T., Shenoy, S. M. & Singer, R. H. Transcriptional pulsing of a developmental gene. Curr. Biol. 16, 1018–1025 (2006).

    Article  CAS  Google Scholar 

  36. Mezo, G., Manea, M., Szabi, I., Vincze, B. & Kovacs, M. New derivatives of GnRH as potential anticancer therapeutic agents. Curr. Med. Chem. 15, 2366–2379 (2008).

    Article  CAS  Google Scholar 

  37. Schacke, H., Docke, W. D. & Asadullah, K. Mechanisms involved in the side effects of glucocorticoids. Pharmacol. Ther. 96, 23–43 (2002).

    Article  CAS  Google Scholar 

  38. Sharp, Z. D. et al. Estrogen-receptor-alpha exchange and chromatin dynamics are ligand- and domain-dependent. J Cell Sci. 119, 4101–4116 (2006).

    Article  CAS  Google Scholar 

  39. Bosisio, D. et al. A hyper-dynamic equilibrium between promoter-bound and nucleoplasmic dimers controls NF-kB-dependent gene activity. EMBO J. 25, 798–810 (2006).

    Article  CAS  Google Scholar 

  40. Hager, G. L. et al. Chromatin dynamics and the evolution of alternate promoter states. Chromosome. Res. 14, 107–116 (2006).

    Article  CAS  Google Scholar 

  41. Muller, W. G., Walker, D., Hager, G. L. & McNally, J. G. Large-scale chromatin decondensation and recondensation regulated by transcription from a natural promoter. J. Cell Biol. 154, 33–48 (2001).

    Article  CAS  Google Scholar 

  42. Vallone, D. et al. Neoplastic transformation of rat thyroid cells requires the junB and fra-1 gene induction which is dependent on the HMGI-C gene product. EMBO J. 16, 5310–5321 (1997).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank T. Karpova and J. McNally for assistance, C. Zeiss Inc. for the opportunity to carry out live cell experiments on a Zeiss Duoscan microscope, D. Shatti for technical assistance with nuclear extract preparation and Y. Kershaw for assistance with the in vivo experiments. This research was supported (in part) by the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research. The animal work was supported by a BBSRC grant (BB/C51297X/1) and a Wellcome Trust Programme Grant (074112/Z/04/Z) to S.L. The qPCR was performed on an Applied Biosystems 7500 System funded by a Wellcome Trust Equipment Grant (075548/Z/04/Z).

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Authors and Affiliations

  1. Laboratory of Receptor Biology and Gene Expression, Center for Cancer Research, National Cancer Institute, NIH, 41 Library Drive, Bethesda, 20892–5055, MD, USA

    Diana A. Stavreva, Malgorzata Wiench, Sam John, Thomas A. Johnson, Ty C. Voss & Gordon L. Hager

  2. Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology, The Dorothy Hodgkin Building, University of Bristol, Whitson Street, Bristol, BS1 3NY, UK

    Becky L. Conway-Campbell, Mervyn A. McKenna, John R. Pooley & Stafford L. Lightman

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  1. Diana A. Stavreva
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Contributions

D.A.S. designed and performed most of the experiments and, together with G.L.H., wrote the initial draft of the manuscript; M.W. and D.A.S. performed transcription experiments, analysed data and participated in manuscript preparation and data interpretation; M.W. also performed all cell-line ChIP experiments; S.J. and T.A.J. performed confirmatory experiments, designed primers for PCR and participated in data interpretation and manuscript revision; T.V. performed some of the single cell analyses, helped with preparation of movies and participated in manuscript revision; B.C.C. and M.M. designed the live animal corticosterone pulsing experiments; J.P. performed surgery, conducted the live animal timecourse experiments and performed ChIP assays; B.CC. performed the TransAM DNA binding assays and prepared figures and text for the in vivo section; M.M. performed the corticosterone EIA and qPCR; S.L. provided data for the corticosterone profile, designed experiments together with G.L.H and D.A.S., provided valuable insights into data interpretation and participated in manuscript preparation; G.L.H. oversaw the project, designed many of the experiments and, together with D.A.S., prepared most figures, coordinated work among collaborators and participated in all stages of manuscript revision.

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Correspondence to Gordon L. Hager.

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Stavreva, D., Wiench, M., John, S. et al. Ultradian hormone stimulation induces glucocorticoid receptor-mediated pulses of gene transcription. Nat Cell Biol 11, 1093–1102 (2009). https://doi.org/10.1038/ncb1922

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