Abstract
In the early embryonic cell cycle, Cdc2–cyclin B functions like an autonomous oscillator, whose robust biochemical rhythm continues even when DNA replication or mitosis is blocked1. At the core of the oscillator is a negative feedback loop; cyclins accumulate and produce active mitotic Cdc2–cyclin B2,3; Cdc2 activates the anaphase-promoting complex (APC); the APC then promotes cyclin degradation and resets Cdc2 to its inactive, interphase state. Cdc2 regulation also involves positive feedback4, with active Cdc2–cyclin B stimulating its activator Cdc25 (refs 5–7) and inactivating its inhibitors Wee1 and Myt1 (refs 8–11). Under the correct circumstances, these positive feedback loops could function as a bistable trigger for mitosis12,13, and oscillators with bistable triggers may be particularly relevant to biological applications such as cell cycle regulation14,15,16,17. Therefore, we examined whether Cdc2 activation is bistable. We confirm that the response of Cdc2 to non-degradable cyclin B is temporally abrupt and switch-like, as would be expected if Cdc2 activation were bistable. We also show that Cdc2 activation exhibits hysteresis, a property of bistable systems with particular relevance to biochemical oscillators. These findings help establish the basic systems-level logic of the mitotic oscillator.
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Acknowledgements
We thank H. Hochegger and T. Hunt for generously providing cyclin B1 antibodies, B. Dunphy for providing a cyclin B1 clone, S. Walter for constructing the Δ65-cyclin B1 clone, T. Guadagno and S. Guadagno for producing and providing the Cdc25 and Wee1 antibodies, M. Hekmat-Nejad for advice on extract preparation, D. Gong for help with experiments, O. Brandmann, B. Novak, W. Sha, J. Sible and J. Tyson for helpful discussions, and K. Cimprich and members of the Ferrell laboratory for comments on the manuscript. This work was supported by the National Institutes of Health (grants GM61276 and GM46383).
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Supplementary information
Supplementary Materials
Part I. Global stability of a two-variable negative fedback system. (PDF 49 kb)
Supplementary Figure 1
Opposing functions in a two-variable negative feedback system. (GIF 11 kb)
Supplementary Materials
Part II. Supplementary figure 2. Stability of 35S-D65-cyclin B1 in M-phase and interphase extracts. 50nM cyclin was added to each type of extract, and the amount of radiolabel remaining was followed as a function of time by SDS-PAGE and autoradiography. (GIF 38 kb)
Supplementary Figure 3
Schematic view of the modeled Cdc2-APC system. (GIF 33 kb)
Supplementary Materials
Part III. Modeling the Cdc2/APC system. (DOC 16 kb)
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Pomerening, J., Sontag, E. & Ferrell, J. Building a cell cycle oscillator: hysteresis and bistability in the activation of Cdc2. Nat Cell Biol 5, 346–351 (2003). https://doi.org/10.1038/ncb954
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DOI: https://doi.org/10.1038/ncb954
