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The Cdk1–APC/C cell cycle oscillator circuit functions as a time-delayed, ultrasensitive switch

Nature Cell Biology volume 15, pages 519–525 (2013)Cite this article

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Abstract

Despite the complexity and variety of biological oscillators, their core design invariably includes an essential negative feedback loop. In the Xenopus laevis embryonic cell cycle oscillator, this loop consists of the kinase cyclin B–Cdk1 and the ubiquitin ligase APC/CCdc20; active Cdk1 activates APC/CCdc20, which then brings about cyclin B degradation and inactivates Cdk1. Here we ask how this negative feedback loop functions quantitatively, with the aim of understanding what mechanisms keep the Cdk1–APC/CCdc20 system from settling into a stable steady state with intermediate levels of Cdk1 and APC/CCdc20 activity. We found that the system operates as a time-delayed, digital switch, with a time lag of ∼ 15 min between Cdk1 and APC/CCdc20 activation and a tremendously high degree of ultrasensitivity (nH≈17). Computational modelling shows how these attributes contribute to the generation of robust, clock-like oscillations. Principles uncovered here may also apply to other activator–repressor oscillators and help in designing robust synthetic clocks.

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Figure 1: Ultrasensitivity in the regulation of securin–CFP destruction by Cdk1.
Figure 2: Ultrasensitive degradation of cyclin A2–YFP and cyclin B1–CFP.
Figure 3: Ultrasensitive cyclin degradation enables limit cycle oscillations.
Figure 4: The EC50 values for APC/CCdc20 activation, Cdc25 phosphorylation and Wee1 phosphorylation are similar.
Figure 5: Time lags in the Xenopus embryonic cell cycle oscillator’s negative feedback loop.

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Acknowledgements

We thank J. Chang and A. Poon for help obtaining purified Δ65-cyclin B1 and Cdk1AF proteins, S. Santos and J. Pomerening for cyclin B1–CFP and cyclin A2–YFP constructs, T. Tsai for sharing his findings on the effects of PD0166285 on Xenopus Wee1A and Myt1, R. Driscoll from the Cimprich laboratory for advice on freezing extracts, and G. Anderson, J. Chang, A. Moskaleva, T. Tsai and the rest of the Ferrell laboratory for scientific discussions and editorial suggestions. We also thank Pfizer for providing PD0166285. Q.Y. is an HHMI Fellow of the Damon Runyon Cancer Research Foundation (DRG-2081-11). This work was supported by the National Institutes of Health grant GM046383.

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Authors and Affiliations

  1. Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, California 94305-5174, USA

    Qiong Yang & James E. Ferrell Jr

  2. Department of Biochemistry, Stanford University School of Medicine, Stanford, California 94305-5174, USA

    James E. Ferrell Jr

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Q.Y. carried out experiments and calculations, analysed data and helped write the paper. J.E.F. carried out calculations, analysed data and helped write the paper.

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Correspondence to James E. Ferrell Jr.

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Yang, Q., Ferrell, J. The Cdk1–APC/C cell cycle oscillator circuit functions as a time-delayed, ultrasensitive switch. Nat Cell Biol 15, 519–525 (2013). https://doi.org/10.1038/ncb2737

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