Positive feedback of G1 cyclins ensures coherent cell cycle entry

Abstract

In budding yeast, Saccharomyces cerevisiae, the Start checkpoint integrates multiple internal and external signals into an all-or-none decision to enter the cell cycle. Here we show that Start behaves like a switch due to systems-level feedback in the regulatory network. In contrast to current models proposing a linear cascade of Start activation, transcriptional positive feedback of the G1 cyclins Cln1 and Cln2 induces the near-simultaneous expression of the 200-gene G1/S regulon. Nuclear Cln2 drives coherent regulon expression, whereas cytoplasmic Cln2 drives efficient budding. Cells with the CLN1 and CLN2 genes deleted frequently arrest as unbudded cells, incurring a large fluctuation-induced fitness penalty due to both the lack of cytoplasmic Cln2 and insufficient G1/S regulon expression. Thus, positive-feedback-amplified expression of Cln1 and Cln2 simultaneously drives robust budding and rapid, coherent regulon expression. A similar G1/S regulatory network in mammalian cells, comprised of non-orthologous genes, suggests either conservation of regulatory architecture or convergent evolution.

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Figure 1: Positive feedback drives the Start of the budding yeast cell cycle.
Figure 2: Cln1 and Cln2 drive coherent expression of the SBF/MBF regulon.
Figure 3: Stochastic unbudded arrest in cln1Δ cln2Δ cells and its modulation by Cln2, Cln3, Whi5, and the mitotic cyclins.
Figure 4: Cln1 and Cln2 are required for rapid phosphorylation and inactivation of the rate-limiting inhibitor Whi5.
Figure 5: Function of nuclear Cln2 and a model for Start regulation by positive feedback.

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Acknowledgements

This work was supported by the National Institute of Health (J.M.S., E.D.S. and F.R.C.), the Burroughs Wellcome Fund (J.M.S.) and the National Science Foundation (E.D.S.). We thank N. Buchler, G. Charvin, B. Drapkin and J. E. Ferrell for conversations; J. Widom and C. Wittenberg for comments on the manuscript; J. M. Bean, B. Timney and J. Robbins for help with strain/plasmid construction; M. Schwab for the plasmid pWS358; B. Futcher for the CLN2-NES and CLN2-NLS plasmids; E. Bi for the pKT355 mCherry tagging plasmid; and M. Tyers for WHI5 phosphorylation site mutant strains and plasmids.

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Correspondence to Jan M. Skotheim.

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Skotheim, J., Di Talia, S., Siggia, E. et al. Positive feedback of G1 cyclins ensures coherent cell cycle entry. Nature 454, 291–296 (2008). https://doi.org/10.1038/nature07118

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