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Autonomous synchronization of the circadian KaiC phosphorylation rhythm

Nature Structural & Molecular Biology volume 14, pages 1084–1088 (2007)Cite this article

Abstract

The cyanobacterial circadian oscillator can be reconstituted in vitro by mixing three purified clock proteins, KaiA, KaiB and KaiC, with ATP. The KaiC phosphorylation rhythm persists for at least 10 days without damping. By mixing oscillatory samples that have different phases and analyzing the dynamics of their phase relationships, we found that the robustness of the KaiC phosphorylation rhythm arises from the rapid synchronization of the phosphorylation state and reaction direction (phosphorylation or dephosphorylation) of KaiC proteins. We further demonstrate that synchronization is tightly linked with KaiC dephosphorylation and is mediated by monomer exchange between KaiC hexamers during the early dephosphorylation phase. This autonomous synchronization mechanism is probably the basis for the resilience of the cyanobacterial circadian system against quantitative fluctuations in clock components during cellular events such as cell growth and division.

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Figure 1: Robust oscillation of KaiC phosphorylation in vitro.
Figure 2: Synchronization of two mixed samples.
Figure 3: Phase relationships between oscillatory mixtures.
Figure 4: The phase of monomer shuffling and its effect on phosphorylation rhythm.
Figure 5: Schematic model of the synchronization machinery of the cyanobacterial circadian clock.

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References

  1. Pittendrigh, C.S. Temporal organization: reflections of a Darwinian clock-watcher. Annu. Rev. Physiol. 55, 16–54 (1993).

    Article  CAS  Google Scholar 

  2. Nakajima, M. et al. Reconstitution of circadian oscillation of cyanobacterial KaiC phosphorylation in vitro. Science 308, 414–415 (2005).

    Article  CAS  Google Scholar 

  3. Kondo, T. et al. Circadian rhythms in rapidly dividing cyanobacteria. Science 275, 224–227 (1997).

    Article  CAS  Google Scholar 

  4. Mihalcescu, I., Hsing, W. & Leibler, S. Resilient circadian oscillator revealed in individual cyanobacteria. Nature 430, 81–85 (2004).

    Article  CAS  Google Scholar 

  5. Amdaoud, M., Vallade, M., Weiss-Schaber, C. & Mihalcescu, I. Cyanobacterial clock, a stable phase oscillator with negligible intercellular coupling. Proc. Natl. Acad. Sci. USA 104, 7051–7056 (2007).

    Article  CAS  Google Scholar 

  6. Kondo, T. et al. Circadian rhythms in prokaryotes: luciferase as a reporter of circadian gene expression in cyanobacteria. Proc. Natl. Acad. Sci. USA 90, 5672–5676 (1993).

    Article  CAS  Google Scholar 

  7. Kageyama, H. et al. Cyanobacterial circadian pacemaker: Kai protein complex dynamics in the KaiC phosphorylation cycle in vitro. Mol. Cell 23, 161–171 (2006).

    Article  CAS  Google Scholar 

  8. Nishiwaki, T. et al. A sequential program of dual phosphorylation of KaiC as a basis for circadian oscillation in cyanobacteria. EMBO J. 26, 4029–4037 (2007).

    Article  CAS  Google Scholar 

  9. van Zon, J.S., Lubensky, D.K., Altena, P.R.H. & ten Wolde, P.R. An allosteric model of circadian KaiC phosphorylation. Proc. Natl. Acad. Sci. USA 104, 7420–7425 (2007).

    Article  CAS  Google Scholar 

  10. Yoda, M., Eguchi, K., Terada, T. & Sasai, M. Monomer-shuffling and allosteric transition in KaiC circadian oscillation. PLoS ONE 2, e408 (2007).

    Article  Google Scholar 

  11. Mori, T. et al. Elucidating the ticking of an in vitro circadian clockwork. PLoS Biol. 5, e93 (2007).

    Article  Google Scholar 

  12. Takigawa-Imamura, H. & Mochizuki, A. Predicting regulation of the phosphorylation cycle of KaiC clock protein using mathematical analysis. J. Biol. Rhythms 21, 405–416 (2006).

    Article  CAS  Google Scholar 

  13. Tomita, J., Nakajima, M., Kondo, T. & Iwasaki, H. No transcription-translation feedback in circadian rhythm of KaiC phosphorylation. Science 307, 251–254 (2005).

    Article  CAS  Google Scholar 

  14. Imai, K., Nishiwaki, T., Kondo, T. & Iwasaki, H. Circadian rhythms in the synthesis and degradation of a master clock protein KaiC in cyanobacteria. J. Biol. Chem. 279, 36534–36539 (2004).

    Article  CAS  Google Scholar 

  15. Elowitz, M.B. & Leibler, S. A synthetic oscillatory network of transcriptional regulators. Nature 403, 335–338 (2000).

    Article  CAS  Google Scholar 

  16. Nagoshi, E. et al. Circadian gene expression in individual fibroblasts: cell-autonomous and self-sustained oscillators pass time to daughter cells. Cell 119, 693–705 (2004).

    Article  CAS  Google Scholar 

  17. Barkai, N. & Leibler, S. Circadian clocks limited by noise. Nature 403, 267–268 (2000).

    Article  CAS  Google Scholar 

  18. Gonze, D., Halloy, J. & Goldbeter, A. Robustness of circadian rhythms with respect to molecular noise. Proc. Natl. Acad. Sci. USA 99, 673–678 (2002).

    Article  CAS  Google Scholar 

  19. Raser, J.M. & O'Shea, E.K. Noise in gene expression: origins, consequences, and control. Science 309, 2010–2013 (2005).

    Article  CAS  Google Scholar 

  20. Elowitz, M.B., Levine, A.J., Siggia, E.D. & Swain, P.S. Stochastic gene expression in a single cell. Science 297, 1183–1186 (2002).

    Article  CAS  Google Scholar 

  21. Rosenfeld, N., Young, J.W., Alon, U., Swain, P.S. & Elowitz, M.B. Gene regulation at the single-cell level. Science 307, 1962–1965 (2005).

    Article  CAS  Google Scholar 

  22. Liu, C., Weaver, D.R., Strogatz, S.H. & Reppert, S.M. Cellular construction of a circadian clock: period determination in the suprachiasmatic nuclei. Cell 91, 855–860 (1997).

    Article  CAS  Google Scholar 

  23. Kitayama, Y., Iwasaki, H., Nishiwaki, T. & Kondo, T. KaiB functions as an attenuator of KaiC phosphorylation in the cyanobacterial circadian clock system. EMBO J. 22, 2127–2134 (2003).

    Article  CAS  Google Scholar 

  24. van den Pol, A.N. in Suprachiasmatic Nucleus: The Mind's Clock (eds. Klein, D.C., Moore, R.Y. & Reppert, S.M.) 17–50 (Oxford University Press, New York, 1991).

    Google Scholar 

Download references

Acknowledgements

We thank H. Kondo and M. Tamura for excellent technical assistance and M. Yamamura (Tokyo Institute of Technology) for comments. This research was supported in part by Grants-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology of Japan (15GS0308 to T.K. and T.O.) and the Japan Society for the Promotion of Science (17370088 to T.O.). H.I. and M.M. were each supported by a Research Fellowship for Young Scientists from the Japan Society for the Promotion of Science (18005539 to H.I. and 17007534 to M.M.).

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  1. Hakuto Kageyama & Masato Nakajima

    Present address: Present addresses: Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, Tokyo 113-0033, Japan (H.K.); Laboratory for Systems Biology, Center for Developmental Biology, RIKEN, 2-2-3 Minatojima-minamimachi, Chuo-ku, Kobe, Hyogo 650-0047, Japan (M.N.).,

Authors and Affiliations

  1. Division of Biological Science, Graduate School of Science, Nagoya University, and Solution Oriented Research for Science and Technology (SORST) Program, Japan Science and Technology Agency (JST), Furo-cho 1, Chikusa-ku, 464-8602, Nagoya, Japan

    Hiroshi Ito, Hakuto Kageyama, Michinori Mutsuda, Masato Nakajima, Tokitaka Oyama & Takao Kondo

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  1. Hiroshi Ito
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Contributions

H.I. and T.O. designed research; H.I., H.K., M.M. and M.N. performed research; H.I., H.K., T.O. and T.K. analyzed data; and H.I., T.O. and T.K. wrote the paper.

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Correspondence to Tokitaka Oyama or Takao Kondo.

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Ito, H., Kageyama, H., Mutsuda, M. et al. Autonomous synchronization of the circadian KaiC phosphorylation rhythm. Nat Struct Mol Biol 14, 1084–1088 (2007). https://doi.org/10.1038/nsmb1312

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