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Abstract	We show that inactivation of the protein kinase Cdk1/Cyclin B (Cdc28/Clb 2 in the budding yeast
Language	English
Publishing date	2023-04-12
Publishing country	United States
Document type	Preprint
DOI	10.21203/rs.3.rs-2787001/v1
Database	MEDical Literature Analysis and Retrieval System OnLINE

Article ; Online: Analog-sensitive Cdk1 as a tool to study mitotic exit: protein phosphatase 1 is required downstream from Cdk1 inactivation in budding yeast.

Keaton, Jason M / Workman, Benjamin G / Xie, Linfeng / Paulson, James R

Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology

2023 Volume 31, Issue 3, Page(s) 27

Abstract: We show that specific inactivation of the protein kinase Cdk1/cyclin B (Cdc28/Clb2) triggers exit from mitosis in the budding yeast Saccharomyces cerevisiae. Cells carrying the allele cdc28-as1, which makes Cdk1 (Cdc28) uniquely sensitive to the ATP ... ...

Abstract	We show that specific inactivation of the protein kinase Cdk1/cyclin B (Cdc28/Clb2) triggers exit from mitosis in the budding yeast Saccharomyces cerevisiae. Cells carrying the allele cdc28-as1, which makes Cdk1 (Cdc28) uniquely sensitive to the ATP analog 1NM-PP1, were arrested with spindle poisons and then treated with 1NM-PP1 to inhibit Cdk1. This caused the cells to leave mitosis and enter G1-phase as shown by initiation of rebudding (without cytokinesis), induction of mating projections ("shmoos") by α-factor, stabilization of Sic1, and degradation of Clb2. It is known that Cdk1 must be inactivated for cells to exit mitosis, but our results show that inactivation of Cdk1 is not only necessary but also sufficient to initiate the transition from mitosis to G1-phase. This result suggests a system in which to test requirements for particular gene products downstream from Cdk1 inactivation, for example, by combining cdc28-as1 with conditional mutations in the genes of interest. Using this approach, we demonstrate that protein phosphatase 1 (PPase1; Glc7 in S. cerevisiae) is required for mitotic exit and reestablishment of interphase following Cdk1 inactivation. This system could be used to test the need for other protein phosphatases downstream from Cdk1 inactivation, such as PPase 2A and Cdc14, and it could be combined with phosphoproteomics to gain information about the substrates that the various phosphatases act upon during mitotic exit.
MeSH term(s)	G1 Phase ; Mitosis ; Protein Phosphatase 1/genetics ; Saccharomyces cerevisiae/genetics ; CDC28 Protein Kinase, S cerevisiae/genetics
Chemical Substances	Protein Phosphatase 1 (EC 3.1.3.16) ; CDC28 Protein Kinase, S cerevisiae (EC 2.7.11.22) ; GLC7 protein, S cerevisiae (EC 3.1.3.16)
Language	English
Publishing date	2023-09-10
Publishing country	Netherlands
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	1161632-5
ISSN	1573-6849 ; 0967-3849
ISSN (online)	1573-6849
ISSN	0967-3849
DOI	10.1007/s10577-023-09736-6
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Exit from Mitosis in Budding Yeast: Protein Phosphatase 1 is Required Downstream from Cdk1 Inactivation.

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