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Updated: Jun 28, 2026

Manipulation and Analysis of Cell Cycle-Dependent Processes in Budding Yeast
Published on: September 26, 2025
Putting the brake on FEAR: Tof2 promotes the biphasic release of Cdc14 phosphatase during mitotic exit
William G Waples1, Charly Chahwan, Marta Ciechonska
1Department of Molecular Genetics, University of Toronto, ON, Canada.
Abstract:
The completion of chromosome segregation during anaphase requires the hypercondensation of the approximately 1-Mb rDNA array, a reaction dependent on condensin and Cdc14 phosphatase. Using systematic genetic screens, we identified 29 novel genetic interactions with budding yeast condensin. Of these, FOB1, CSM1, LRS4, and TOF2 were required for the mitotic condensation of the tandem rDNA array localized on chromosome XII. Interestingly, whereas Fob1 and the monopolin subunits Csm1 and Lrs4 function in rDNA condensation throughout M phase, Tof2 was only required during anaphase. We show that Tof2, which shares homology with the Cdc14 inhibitor Net1/Cfi1, interacts with Cdc14 phosphatase and its deletion suppresses defects in mitotic exit network (MEN) components. Consistent with these genetic data, the onset of Cdc14 release from the nucleolus was similar in TOF2 and tof2Delta cells; however, the magnitude of the release was dramatically increased in the absence of Tof2, even when the MEN pathway was compromised. These data support a model whereby Tof2 coordinates the biphasic release of Cdc14 during anaphase by restraining a population of Cdc14 in the nucleolus after activation of the Cdc14 early anaphase release (FEAR) network, for subsequent release by the MEN.
Insights
Tof2 restrains Cdc14 phosphatase release from the nucleolus during anaphase, ensuring proper chromosome segregation. This regulation is crucial for coordinating the biphasic release of Cdc14, essential for cell division.
Area of Science:
- Cell Biology
- Molecular Biology
- Genetics
Background:
- Chromosome segregation requires rDNA array hypercondensation, dependent on condensin and Cdc14 phosphatase.
- Understanding the regulation of rDNA condensation and Cdc14 phosphatase activity is crucial for cell cycle progression.
Purpose of the Study:
- To identify novel genetic interactions involved in budding yeast condensin function.
- To elucidate the role of specific genes, including TOF2, in rDNA condensation and chromosome segregation.
- To understand the regulatory mechanism of Cdc14 phosphatase release during anaphase.
Main Methods:
- Systematic genetic screens to identify interactions with budding yeast condensin.
- Analysis of gene deletion mutants (e.g., tof2Δ) to assess their role in rDNA condensation.
- Investigation of protein interactions and localization (e.g., Cdc14 release from the nucleolus).
Main Results:
- Identified FOB1, CSM1, LRS4, and TOF2 as essential for mitotic rDNA condensation.
- Demonstrated that Tof2, unlike Fob1, Csm1, and Lrs4, is specifically required during anaphase.
- Showed that Tof2 interacts with Cdc14 phosphatase and its absence enhances Cdc14 release, even when the mitotic exit network (MEN) is compromised.
Conclusions:
- Tof2 plays a critical role in coordinating the biphasic release of Cdc14 phosphatase during anaphase.
- Tof2 restrains Cdc14 in the nucleolus after FEAR network activation, allowing for subsequent MEN-mediated release.
- This regulation by Tof2 is vital for timely and accurate chromosome segregation.
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