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Scaffold-mediated symmetry breaking by Cdc42p

Nature Cell Biology volume 5, pages 1062–1070 (2003)Cite this article

Abstract

Cell polarization generally occurs along a single well-defined axis that is frequently determined by environmental cues such as chemoattractant gradients or cell–cell contacts, but polarization can also occur spontaneously in the apparent absence of such cues, through a process called symmetry breaking1,2,3,4,5. In Saccharomyces cerevisiae, cells are born with positional landmarks that mark the poles of the cell and guide subsequent polarization and bud emergence to those sites, but cells lacking such landmarks polarize towards a random cortical site and proliferate normally6. The landmarks employ a Ras-family GTPase, Rsr1p7,8,9, to communicate with the conserved Rho-family GTPase Cdc42p, which is itself polarized and essential for cytoskeletal polarization10,11. We found that yeast Cdc42p was effectively polarized to a single random cortical site even in the combined absence of landmarks, microtubules and microfilaments. Among a panel of Cdc42p effectors and interacting proteins, we found that the scaffold protein Bem1p was uniquely required for this symmetry-breaking behaviour. Moreover, polarization was dependent on GTP hydrolysis by Cdc42p, suggesting that assembly of a polarization site involves cycling of Cdc42p between GTP- and GDP-bound forms, rather than functioning as a simple on/off switch.

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Figure 1: Landmark-independent Cdc42p polarization does not require polymerized actin or tubulin.
Figure 2: Landmark-independent Cdc42p polarization requires Bem1p.
Figure 3: Cdc42p function requires GTP hydrolysis.
Figure 4: Spontaneous Cdc42pQ61L polarization only occurs after overexpression to toxic levels.
Figure 5: Cdc42p function in symmetry breaking and polarity establishment.

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Acknowledgements

We wish to thank Y. Matsui and T. Ito for the generous gift of plasmids, P. Brennwald for the generous gift of affinity-purified anti-Cdc42p and anti-Bem1p antibodies, and L. Schenkman and J. Pringle for the generous gift of strains and primers. We also wish to thank S. Kornbluth, D. Kiehart and S. Haase for critical reading of the manuscript, S. Margolis and D. Colón-Ramos, and members of the Pringle and Lew labs for stimulating discussions. This work was supported by a grant GM62300 from the National Institutes of Health/National Institute of General Medical Sciences to D.J.L. J.E.I. was supported by a Predoctoral Fellowship DAMD17-01-1-0231 from the Department of Defense Breast Cancer Research Program.

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  1. Department of Pharmacology and Cancer Biology Duke University Medical Center, Durham, 27710, NC, USA

    Javier E. Irazoqui, Amy S. Gladfelter & Daniel J. Lew

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Correspondence to Daniel J. Lew.

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Irazoqui, J., Gladfelter, A. & Lew, D. Scaffold-mediated symmetry breaking by Cdc42p. Nat Cell Biol 5, 1062–1070 (2003). https://doi.org/10.1038/ncb1068

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