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Stabilization of microtubule dynamics at anaphase onset promotes chromosome segregation

Nature volume 433, pages 171–176 (2005)Cite this article

Abstract

Microtubules of the mitotic spindle form the structural basis for chromosome segregation. In metaphase, microtubules show high dynamic instability, which is thought to aid the ‘search and capture’ of chromosomes for bipolar alignment on the spindle. Microtubules suddenly become more stable at the onset of anaphase, but how this change in microtubule behaviour is regulated and how important it is for the ensuing chromosome segregation are unknown1,2,3,4. Here we show that in the budding yeast Saccharomyces cerevisiae, activation of the phosphatase Cdc14 at anaphase onset is both necessary and sufficient for silencing microtubule dynamics. Cdc14 is activated by separase, the protease that triggers sister chromatid separation, linking the onset of anaphase to microtubule stabilization5,6. If sister chromatids separate in the absence of Cdc14 activity, microtubules maintain high dynamic instability; this correlates with defects in both the movement of chromosomes to the spindle poles (anaphase A) and the elongation of the anaphase spindle (anaphase B). Cdc14 promotes localization of microtubule-stabilizing proteins to the anaphase spindle, and dephosphorylation of the kinetochore component Ask1 contributes to both the silencing of microtubule turnover and successful anaphase A.

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Figure 1: Separase stabilizes microtubule dynamics at anaphase onset.
Figure 2: Cdc14 phosphatase regulates microtubule dynamics in mitosis.
Figure 3: Cdc14 activation is required for successful anaphase A.
Figure 4: Anaphase B defects during TEV protease-triggered anaphase.
Figure 5: Proteins involved in the Cdc14 response.

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Acknowledgements

We thank S. Elledge, J. Kilmartin and A. Straight for reagents, R. Carazo-Salas and A. Nicol for advice on microscopy, J. Cau, T. Davis, A. Hyman, T. Toda and all members of our laboratory for helpful discussions and critical reading of the manuscript, and in particular M. Sullivan for help at the outset of this study.

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Authors and Affiliations

  1. Chromosome Segregation Laboratory, Cancer Research UK London Research Institute, Lincoln's Inn Fields Laboratories, 44 Lincoln's Inn Fields, WC2A 3PX, London, UK

    Toru Higuchi & Frank Uhlmann

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  1. Toru Higuchi
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  2. Frank Uhlmann
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Correspondence to Frank Uhlmann.

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The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Figure S1

Summary of microtubule dynamics measurements. (PDF 389 kb)

Supplementary Figure S2

Cdc14 rescues spindle stability during TEV protease-triggered anaphase. (PDF 795 kb)

Supplementary Figure S3

Ectopic Cdc14 in metaphase reduces centromere oscillations. (PDF 182 kb)

Supplementary Figure S4

Rescue of spindle stability in the absence of Cdc14 by deletion of Kip3. (PDF 187 kb)

Supplementary Movie 1

Spindle elongation during separase-triggered anaphase. (MPG 339 kb)

Supplementary Movie 2

Spindle elongation during TEV protease-triggered anaphase, example 1. (MPG 251 kb)

Supplementary Movie 3

Spindle elongation during TEV protease-triggered anaphase, example 2 (MPG 207 kb)

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Higuchi, T., Uhlmann, F. Stabilization of microtubule dynamics at anaphase onset promotes chromosome segregation. Nature 433, 171–176 (2005). https://doi.org/10.1038/nature03240

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