Mitotic Exit in the Absence of Separase Activity

doi:10.1091/MBC.E08-10-1042

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Mitotic Exit in the Absence of Separase Activity

Journal Article•DOI•

Mitotic Exit in the Absence of Separase Activity

Ying Lu¹, Frederick R. Cross¹•Institutions (1)

Rockefeller University¹

01 Mar 2009-Molecular Biology of the Cell (American Society for Cell Biology)-Vol. 20, Iss: 5, pp 1576-1591

TL;DR: The first quantitative measure for Cdc14 release based on colocalization with the Net1 nucleolar anchor is defined, indicating efficient CDC14 release upon MEN activation; release driven by Esp1 in the absence of microtubules was inefficient and incapable of driving ME.

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Abstract: In budding yeast, three interdigitated pathways regulate mitotic exit (ME): mitotic cyclin–cyclin-dependent kinase (Cdk) inactivation; the Cdc14 early anaphase release (FEAR) network, including a n...

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Journal Article•DOI•

Phosphatases: providing safe passage through mitotic exit

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Claudia Wurzenberger¹, Daniel W. Gerlich¹•Institutions (1)

ETH Zurich¹

01 Aug 2011-Nature Reviews Molecular Cell Biology

TL;DR: This work has shown that the key mitotic exit phosphatase in budding yeast, Cdc14, is now well understood, and in animal cells, it is now emerging that mitoticexit relies on distinct regulatory networks, including the protein phosphatases PP1 and PP2A.

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Abstract: The mitosis-to-interphase transition involves dramatic cellular reorganization from a state that supports chromosome segregation to a state that complies with all functions of an interphase cell. This process, termed mitotic exit, depends on the removal of mitotic phosphorylations from a broad range of substrates. Mitotic exit regulation involves inactivation of mitotic kinases and activation of counteracting protein phosphatases. The key mitotic exit phosphatase in budding yeast, Cdc14, is now well understood. By contrast, in animal cells, it is now emerging that mitotic exit relies on distinct regulatory networks, including the protein phosphatases PP1 and PP2A.

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294 citations

Journal Article•DOI•

Periodic cyclin-Cdk activity entrains an autonomous Cdc14 release oscillator.

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Ying Lu¹, Frederick R. Cross¹•Institutions (1)

Rockefeller University¹

16 Apr 2010-Cell

TL;DR: An intrinsically oscillatory module controlling nucleolar release and resequestration of the Cdc14 phosphatase is demonstrated, which is essential for mitotic exit in budding yeast and suggests that the intrinsically autonomous CDC14 release cycles are locked at once-per-cell-cycle through entrainment by the Cdk oscillator in wild-type cells.

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97 citations

Cites methods from "Mitotic Exit in the Absence of Sepa..."

...Cdc14 release was quantified as the coefficient of variation (CV, standard deviation divided by mean) of Cdc14-YFP pixel intensities (mother and bud treated separately), divided by the Net1-mCherry CV....
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...We determined the response of the Cdc14 release cycle to fixed cyclin-Cdk levels (Drapkin et al., 2009), using a quantitative, single cell measurement for Cdc14 localization based on variation of cellular Cdc14-YFP pixel intensities, standardized to variation of nucleolar Net1-mCherry (Lu and Cross, 2009) (Experimental Procedures; Figure 1A)....
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...…cycle to fixed cyclin-Cdk levels (Drapkin et al., 2009), using a quantitative, single cell measurement for Cdc14 localization based on variation of cellular Cdc14-YFP pixel intensities, standardized to variation of nucleolar Net1-mCherry (Lu and Cross, 2009) (Experimental Procedures; Figure 1A)....
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..., 2009), using a quantitative, single cell measurement for Cdc14 localization based on variation of cellular Cdc14-YFP pixel intensities, standardized to variation of nucleolar Net1-mCherry (Lu and Cross, 2009) (Experimental Procedures; Figure 1A)....
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Journal Article•DOI•

Inhibition of Cdc42 during mitotic exit is required for cytokinesis

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Benjamin D. Atkins¹, Satoshi Yoshida², Satoshi Yoshida¹, Koji Saito³, Chi-Fang Wu³, Daniel J. Lew³, David Pellman¹, David Pellman⁴ - Show less +4 more•Institutions (4)

Harvard University¹, Brandeis University², Duke University³, Howard Hughes Medical Institute⁴

22 Jul 2013-Journal of Cell Biology

TL;DR: A decrease in Cdc42 activation during mitotic exit is necessary to allow localization of key cytokinesis regulators and proper septum formation.

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Abstract: The role of Cdc42 and its regulation during cytokinesis is not well understood. Using biochemical and imaging approaches in budding yeast, we demonstrate that Cdc42 activation peaks during the G1/S transition and during anaphase but drops during mitotic exit and cytokinesis. Cdc5/Polo kinase is an important upstream cell cycle regulator that suppresses Cdc42 activity. Failure to down-regulate Cdc42 during mitotic exit impairs the normal localization of key cytokinesis regulators-Iqg1 and Inn1-at the division site, and results in an abnormal septum. The effects of Cdc42 hyperactivation are largely mediated by the Cdc42 effector p21-activated kinase Ste20. Inhibition of Cdc42 and related Rho guanosine triphosphatases may be a general feature of cytokinesis in eukaryotes.

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79 citations

Journal Article•DOI•

Global analysis of Cdc14 phosphatase reveals diverse roles in mitotic processes.

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Joanna Bloom¹, Joanna Bloom², Ileana M. Cristea², Andrea L. Procko², Veronica Lubkov², Brian T. Chait², Michael Snyder¹, Frederick R. Cross² - Show less +4 more•Institutions (2)

Yale University¹, Rockefeller University²

18 Feb 2011-Journal of Biological Chemistry

TL;DR: The findings suggest the dephosphorylation of the formins may be important for their observed localization change during exit from mitosis and indicate that Cdc14 targets proteins involved in wide-ranging mitotic events.

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79 citations

Journal Article•DOI•

From START to FINISH: computational analysis of cell cycle control in budding yeast.

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Pavel Kraikivski¹, Katherine C. Chen¹, Teeraphan Laomettachit², T. M. Murali¹, John J. Tyson¹ - Show less +1 more•Institutions (2)

Virginia Tech¹, King Mongkut's University of Technology Thonburi²

10 Dec 2015

TL;DR: This work crafted a new mathematical model of cell cycle progression in yeast that exploits a natural separation of time scales in the cell cycle control network to construct a system of differential-algebraic equations for protein synthesis and degradation, post-translational modifications, and rapid formation and dissociation of multimeric complexes.

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Abstract: In the cell division cycle of budding yeast, START refers to a set of tightly linked events that prepare a cell for budding and DNA replication, and FINISH denotes the interrelated events by which the cell exits from mitosis and divides into mother and daughter cells. On the basis of recent progress made by molecular biologists in characterizing the genes and proteins that control START and FINISH, we crafted a new mathematical model of cell cycle progression in yeast. Our model exploits a natural separation of time scales in the cell cycle control network to construct a system of differential-algebraic equations for protein synthesis and degradation, post-translational modifications, and rapid formation and dissociation of multimeric complexes. The model provides a unified account of the observed phenotypes of 257 mutant yeast strains (98% of the 263 strains in the data set used to constrain the model). We then use the model to predict the phenotypes of 30 novel combinations of mutant alleles. Our comprehensive model of the molecular events controlling cell cycle progression in budding yeast has both explanatory and predictive power. Future experimental tests of the model's predictions will be useful to refine the underlying molecular mechanism, to constrain the adjustable parameters of the model, and to provide new insights into how the cell division cycle is regulated in budding yeast.

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69 citations

Additional excerpts

...In this case, anaphase proceeds normally, activating the MEN while the FEAR pathway remains blocked.(42) In agreement with Lu and Cross,(42) our simulations (Figure 3) show that MEN activity is absolutely needed for mitotic exit, and FEAR is dispensable....
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Journal Article•DOI•

Separase, polo kinase, the kinetochore protein Slk19, and Spo12 function in a network that controls Cdc14 localization during early anaphase.

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Frank Stegmeier¹, Rosella Visintin¹, Angelika Amon¹•Institutions (1)

Massachusetts Institute of Technology¹

25 Jan 2002-Cell

TL;DR: A regulatory network, the FEAR (Cdc fourteen early anaphase release) network that promotes Cdc14 release from the nucleolus during earlyAnaphase is identified and it is proposed that one function of CDC14 released by the Fear network is to stimulate MEN activity.

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476 citations

"Mitotic Exit in the Absence of Sepa..." refers background or result in this paper

...…Net1, contributing to early release of Cdc14 (Azzam et al., 2004); in addition, the MEN may be inhibited by Clb-Cdk, at least at very high levels (Jaspersen and Morgan, 2000; Stegmeier et al., 2002), and Cdc14 released by the MEN promotes Clb– Cdk inactivation (Stegmeier and Amon, 2004)....
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...Blocking sister separation with Scc1-RRDD in the presence of active Esp1 causes a delay in ME estimated between 20 and 60 min, depending on the assays for ME and/or on the exact experimental conditions (Uhlmann et al., 2000; Stegmeier et al., 2002)....
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...The minor delay of ME in the GAL1-SIC1-4A GAL1-TEV/SCC1-TEV cdc20-blocked cells compared with release by Cdc20 reactivation is consistent with the lack of FEAR network activity due to lack of Esp1 activity (Stegmeier et al., 2002)....
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...Cdc15 may be phosphorylated by cyclin (Clb)-Cdk, potentially leading to inhibition of the MEN (Jaspersen and Morgan, 2000); despite this, high Clb-Cdk does not block Cdc14 release from the nucleolus (Stegmeier et al., 2002)....
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...15–20 min (Stegmeier et al., 2002), shorter than our estimate of the time required for ME in the presence of active Esp1, but without cohesin cleavage....
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Journal Article•DOI•

Closing Mitosis: The Functions of the Cdc14 Phosphatase and Its Regulation

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Frank Stegmeier¹, Angelika Amon¹•Institutions (1)

Massachusetts Institute of Technology¹

29 Nov 2004-Annual Review of Genetics

TL;DR: The functions of Cdc14 are reviewed and how this phosphatase is regulated to accomplish the coupling of mitotic processes.

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Abstract: ▪ Abstract Completion of the cell cycle requires the temporal and spatial coordination of chromosome segregation with mitotic spindle disassembly and cytokinesis. In budding yeast, the protein phosphatase Cdc14 is a key regulator of these late mitotic events. Here, we review the functions of Cdc14 and how this phosphatase is regulated to accomplish the coupling of mitotic processes. We also discuss the function and regulation of Cdc14 in other eukaryotes, emphasizing conserved features.

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449 citations

"Mitotic Exit in the Absence of Sepa..." refers background in this paper

...…Net1, contributing to early release of Cdc14 (Azzam et al., 2004); in addition, the MEN may be inhibited by Clb-Cdk, at least at very high levels (Jaspersen and Morgan, 2000; Stegmeier et al., 2002), and Cdc14 released by the MEN promotes Clb– Cdk inactivation (Stegmeier and Amon, 2004)....
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...Cdc14 is essential for ME, but it cannot promote ME until it is released from its nucleolar anchor Net1 (Stegmeier and Amon, 2004)....
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...Spindle elongation could promote ME by driving the daughter spindle pole into proximity to the bud cortex, activating the MEN (Yeh et al., 1995; Bardin et al., 2000; Molk et al., 2004; Stegmeier and Amon, 2004)....
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..., 2002), and Cdc14 released by the MEN promotes Clb– Cdk inactivation (Stegmeier and Amon, 2004)....
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...It is well established that the activity of Cdc14 is regulated by its localization in the nucleolus, where it is stably bound to its inhibitor Net1 and also sequestered from many potential dephosphorylation targets (Stegmeier and Amon, 2004)....
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Journal Article•DOI•

Inhibitory phosphorylation of the APC regulator Hct1 is controlled by the kinase Cdc28 and the phosphatase Cdc14.

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Sue L. Jaspersen¹, Julia F. Charles¹, David O. Morgan¹•Institutions (1)

University of California, San Francisco¹

11 Mar 1999-Current Biology

TL;DR: It is found that Hct1 was phosphorylated in vivo at multiple CDK consensus sites during cell cycle stages when activity of the cyclin-dependent kinase Cdc28 is high and APC activity is low, and phosphorylation abolished the ability of HCT1 to activate the APC in vitro.

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439 citations

"Mitotic Exit in the Absence of Sepa..." refers background in this paper

...…degradation, and all other aspects of ME scored, were dependent on an intact spindle, the MEN, and Cdh1, in Esp1 overexpressors, suggesting that full cdc20 bypass by overexpressed Esp1 requires Clb–Cdk inactivation via Cdh1 activation through the MEN (Visintin et al., 1998; Jaspersen et al., 1999)....
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...MEN activation promotes highly efficient Cdc14 release, which can activate Cdh1 by dephosphorylation (Visintin et al., 1998; Jaspersen et al., 1999), leading to Clb degradation....
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...Clb2 degradation, and all other aspects of ME scored, were dependent on an intact spindle, the MEN, and Cdh1, in Esp1 overexpressors, suggesting that full cdc20 bypass by overexpressed Esp1 requires Clb–Cdk inactivation via Cdh1 activation through the MEN (Visintin et al., 1998; Jaspersen et al., 1999)....
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Journal Article•DOI•

Involvement of an Actomyosin Contractile Ring in Saccharomyces cerevisiae Cytokinesis

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Erfei Bi¹, Paul S. Maddox, Daniel J. Lew², Edward D. Salmon, John N. McMillan², Elaine Yeh, John R. Pringle¹ - Show less +3 more•Institutions (2)

University of North Carolina at Chapel Hill¹, Duke University²

07 Sep 1998-Journal of Cell Biology

TL;DR: The contractile actomyosin ring is not essential for cytokinesis in S. cerevisiae, and in its absence, cytokineses can still be completed by a process that appears to require septin function and to be facilitated by F-actin.

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Abstract: In Saccharomyces cerevisiae, the mother cell and bud are connected by a narrow neck. The mechanism by which this neck is closed during cytokinesis has been unclear. Here we report on the role of a contractile actomyosin ring in this process. Myo1p (the only type II myosin in S. cerevisiae) forms a ring at the presumptive bud site shortly before bud emergence. Myo1p ring formation depends on the septins but not on F-actin, and preexisting Myo1p rings are stable when F-actin is depolymerized. The Myo1p ring remains in the mother–bud neck until the end of anaphase, when a ring of F-actin forms in association with it. The actomyosin ring then contracts to a point and disappears. In the absence of F-actin, the Myo1p ring does not contract. After ring contraction, cortical actin patches congregate at the mother–bud neck, and septum formation and cell separation rapidly ensue. Strains deleted for MYO1 are viable; they fail to form the actin ring but show apparently normal congregation of actin patches at the neck. Some myo1Δ strains divide nearly as efficiently as wild type; other myo1Δ strains divide less efficiently, but it is unclear whether the primary defect is in cytokinesis, septum formation, or cell separation. Even cells lacking F-actin can divide, although in this case division is considerably delayed. Thus, the contractile actomyosin ring is not essential for cytokinesis in S. cerevisiae. In its absence, cytokinesis can still be completed by a process (possibly localized cell–wall synthesis leading to septum formation) that appears to require septin function and to be facilitated by F-actin.

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420 citations

"Mitotic Exit in the Absence of Sepa..." refers background or methods in this paper

...We included a Myo1–green fluorescent protein (GFP) marker (Bi et al., 1998) to allow measurement of the time between budding and cytokinesis (determined by Myo1 ring disappearance), and observed a delay of 0.5 h due to SCC1-RRDD expression....
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...We included a Myo1–green fluorescent protein (GFP) marker (Bi et al., 1998) to allow measurement of the time between budding and cytokinesis (determined by Myo1 ring disappearance), and observed a delay of 0....
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...Myo1-mCherry forms a ring at the bud neck which disappears at cytokinesis (Bi et al., 1998)....
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Journal Article•DOI•

CLB5: a novel B cyclin from budding yeast with a role in S phase.

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Charles B. Epstein¹, Frederick R. Cross•Institutions (1)

Rockefeller University¹

01 Sep 1992-Genes & Development

TL;DR: Observations are suggestive of partial functional redundancy between CLB5 and CLN genes, which is the only yeast cyclin whose deletion lengthens S phase and may have some role in promoting the G1/S transition.

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Abstract: Budding yeast strains have three CLN genes, which have limited cyclin homology. At least one of the three is required for cell cycle START. Four B cyclins are known in yeast; two have been shown to function in mitosis. We have discovered a fifth B-cyclin gene, called CLBS, which when cloned on a CEN plasmid can rescue strains deleted for all three CLN genes. CLB5 transcript abundance peaks in G~, coincident with the CLN2 transcript but earlier than the CLB2 transcript. CLB5 deletion does not cause lethality, either alone or in combination with other CLN or CLB deletions. However, strains deleted for CLB5 require more time to complete S phase, suggesting that CLB5 promotes some step in DNA synthesis. CLB5 is the only yeast cyclin whose deletion lengthens S phase. CLB5 may also have some role in promoting the GI/S transition, because clnl cln2 strains require both CLN3 and CLB5 for viability on glycerol media and clnl,2,3- strains require CLB5 for rescue by the Drosophila melanogaster cdc2 gene. In conjunction with clnl,2,3- rescue by CLB5 overexpression and the coincident transcriptional regulation of CLB5 and CLN2, these observations are suggestive of partial functional redundancy between CLB5 and CLN genes.

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391 citations