Orchestrating anaphase and mitotic exit: separase cleavage and localization of Slk19

Abstract

Anaphase in budding yeast is triggered by cleavage of the central subunit, Scc1, of the chromosomal cohesin complex by the protease separase. Here we show that separase also cleaves the kinetochore-associated protein Slk19 at anaphase onset. Separase activity is also required for the proper localization of a stable Slk19 cleavage product to the spindle midzone in anaphase. The cleavage and localization of Slk19 are necessary to stabilize the anaphase spindle, and we show that a stable spindle is a prerequisite for timely exit from mitosis. This demonstrates the cleavage of targets other than cohesin by separase in the orchestration of high-fidelity anaphase.

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Figure 1: Slk19 is a cleavage target of separase.
Figure 2: Phenotype of uncleavable Slk19.
Figure 3: The Slk19 cleavage product is required for spindle stability in anaphase.
Figure 4: Delay of mitotic exit after spindle breakdown.
Figure 5: Cleavage of Slk19 by TEV protease.
Figure 6: Separase activity in anaphase is required for Slk19 and Ase1 localization.

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Acknowledgements

We thank J. Kilmartin for his gift of the anti-Tub4 antibody, K. Sawin for the anti-GFP antibody, K. Nasmyth for yeast strains and for his support and encouragement, and J. Diffley and T. Toda for critical comments on the manuscript.

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Correspondence to Frank Uhlmann.

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