Turkish Journal of Biology
Abstract
Background/aim: The conserved phosphatase Cdc14 facilitates mitotic exit in budding yeast by counteracting mitotic Cyclin Dependent Kinase activity. Cdc14 is kept in the nucleolus until anaphase-onset, when its released transiently into the nucleoplasm. In late-anaphase, Cdc14 is fully released into the cytoplasm upon activation of the Mitotic Exit Network (MEN) to trigger mitotic exit. Cdc14 also localizes to yeast Spindle Pole Bodies (SPB) in anaphase and dephosphorylates key targets that resides at SPBs to license SPB duplication and prime the MEN. Protein Phosphatase 1 (Glc7) with its regulatory-subunit Bud14 is another phosphatase that plays a key role in spatiotemporal control of mitotic exit. Here, we investigated regulation of Cdc14 localization by Bud14-Glc7.Materials and methods : We used fluorescence microscopy to analyze Cdc14 localization in BUD14 wildtype and BUD14 knockout cells (bud14∆) as well as in cells expressing a mutant allele of BUD14 (bud14-F379A) that cannot bind Glc7. We also utilized yeast two-hybrid (Y2H) to examine interaction of Bud14 with Cdc14.Results: We show that Cdc14 stays longer at the SPBs in bud14∆ and bud14-F379A compared to the wildtype cells. This effect is limited to the SPB that migrated to the daughter cell (dSPB). Cdc14 localizes to both SPBs shortly after anaphase-onset. In mid-to-late anaphase, levels of Cdc14 increases at the dSPB both in wildtype and bud14∆ cells. With mitotic exit, Cdc14 disappears from dSPB in wildtype but not in bud14∆ cells. Accordingly, 50% of bud14∆ cells in G1 have Cdc14 at SPBs. We further find that Cdc14 localization at the dSPB is largely, but not entirely, dependent on Bfa1 in bud14∆ cells. In addition, we show that Bud14 interacts with Cdc14 in Y2H.Conclusion: Our results suggest that Glc7-Bud14 is part of a mechanism that promote Cdc14 disappearance from the dSPB.
DOI
10.55730/1300-0152.2707
Keywords
Bud14, Cdc14, Mitotic Exit Network, Protein Phosphatase 1, Spindle Pole Body
First Page
308
Last Page
318
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.
Recommended Citation
KIRDÖK, İDİL and KOCA ÇAYDAŞI, AYŞE
(2024)
"Disappearance of Cdc14 from the daughter spindle pole body requires Glc7-Bud14,"
Turkish Journal of Biology: Vol. 48:
No.
5, Article 5.
https://doi.org/10.55730/1300-0152.2707
Available at:
https://journals.tubitak.gov.tr/biology/vol48/iss5/5
