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A non-proteolytic function of separase links the onset of anaphase to mitotic exit

Nature Cell Biology volume 5, pages 249–254 (2003)Cite this article

Abstract

Separase is a protease that triggers chromosome segregation at anaphase onset by cleaving cohesin, the chromosomal protein complex responsible for sister chromatid cohesion1,2. After anaphase, cells exit from mitosis; that is, they complete downregulation of cyclin-dependent kinase activity, undergo cytokinesis and enter G1 of the next cell cycle. Here we show that separase activation at the onset of anaphase is sufficient to promote release from the nucleolus and activation of the budding yeast phosphatase, Cdc14, a key step in mitotic exit3,4,5. The ability of separase to activate Cdc14 is independent of its protease function but may involve promoting phosphorylation of the Cdc14 inhibitor Net1. This novel separase function is coregulated with its proteolytic activity by the separase inhibitor securin. This helps to explain the coupling of anaphase and mitotic exit — after securin degradation at anaphase onset, separase cleaves cohesin to trigger chromosome segregation and concurrently uses a non-proteolytic mechanism to initiate mitotic exit.

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Figure 1: Separase promotes mitotic exit.
Figure 2: Analysis of nucleolar Cdc14 release by separase.
Figure 3: Release of Cdc14 is independent of separase protease activity.
Figure 4: Interactions of separase during mitotic exit.
Figure 5: A model for anaphase and mitotic exit.

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Acknowledgements

We wish to thank J. Aris, O. Cohen-Fix, L. Johnston, K. Sawin, and A. Toh-e for reagents, S. Weitzer for cloning Polo, A. Amon, J. Cooper, J. Hayles, M. Pardo, E. Roldan, T. Toda and all members of the laboratory for discussions and comments on the manuscript.

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

    Matt Sullivan & Frank Uhlmann

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Correspondence to Frank Uhlmann.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Figures and References

Supplementary Figure S1. Separase-induced rebudding depends upon Cdc14 activity. (PDF 176 kb)

Supplementary Figure S2. Separase expression leads to dephosphorylation of Cdc15 and accumulation of Sic1.

Supplementary Figure S3. Overexpression of neither Slk19 nor Spo12 in metaphase arrested cells induces nucleolar release of Cdc14.

Supplementary Figure S4. Slk19 cleavage is not required for Cdc14 nucleolar release.

Supplementary Figure S5. The localisation of separase in anaphase depends on Slk19

Supplementary Figure S6. Analysis of Polo mediated nucleolar release of Cdc14.

Supplementary Figure S7. Comparison of Cdc14 release by Polo or separase expression suggests that Polo is less efficient in releasing Cdc14.

Supplementary Figure S8. Deletion of Slk19 delays mitotic exit independently of anaphase spindle destabilisation.

Supplementary references

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Sullivan, M., Uhlmann, F. A non-proteolytic function of separase links the onset of anaphase to mitotic exit. Nat Cell Biol 5, 249–254 (2003). https://doi.org/10.1038/ncb940

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