Molecular control of the Wee1 regulatory pathway by the SAD kinase Cdr2

Guzmán-Vendrell, Mercè; Rincón, Sergio A.; Dingli, Florent; Loew, Damarys; Paoletti, Anne

doi:10.1242/jcs.173146

Cited by 17 publications

(25 citation statements)

References 81 publications

(135 reference statements)

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“…To begin investigating how Wee1 is regulated by nodes, we tested its physical association with Cdr2. In a yeast two-hybrid assay, Wee1 was previously shown to interact with the Cdr2 kinase domain, and this interaction was lost in the catalytically inactive cdr2(E177A) mutant (Guzman-Vendrell et al, 2015). We confirmed this interaction, and then used it to determine the domain of Wee1 critical for Cdr2 interaction and for localization to nodes.…”

Section: Wee1 Localization To Nodes Requires Cdr2 Kinase Activity Andsupporting

confidence: 53%

“…This mutant lacks the C-terminal KA1 domain that binds to both lipids and to Cdr1, but it retains the Cdr2 kinase and UBA domains that are required for catalytic activity. As a result, Cdr2∆C does not bind the cortex or form nodes, and instead localizes diffusely to the cytoplasm and nucleus ( Figure 1B) (Deng et al, 2014;Guzman-Vendrell et al, 2015;Moravcevic et al, 2010). We observed loss of Wee1 phosphorylation in the cdr2∆C mutant similar to a cdr2∆ mutant ( Figure 1C).…”

Section: Results and Discussion: Nodes Are Stable Complexes That Prommentioning

confidence: 71%

“…These results show that Cdr1 and Cdr2 regulate phosphorylation of Wee1 in cells, and point to a key role for Cdr2 in this process. Cdr2 assembles into stationary nodes at the plasma membrane during G2, and recruits Cdr1 to these sites (Guzman-Vendrell et al, 2015;Martin and Berthelot-Grosjean, 2009;Morrell et al, 2004;Moseley et al, 2009). To test if assembly of Cdr2 and Cdr1 into nodes is important for Wee1 phosphorylation in cells, we blotted against Wee1 in cdr2∆C cells.…”

Section: Results and Discussion: Nodes Are Stable Complexes That Prommentioning

confidence: 99%

“…Pom1 directly phosphorylates Cdr2 to inhibit mitotic entry through Wee1. This modification inhibits node assembly and prevents the activating kinase Ssp1 from phosphorylating the Cdr2 activation loop Deng et al, 2014;Guzman-Vendrell et al, 2015;Kettenbach et al, 2015;Moseley et al, 2009;Rincon et al, 2014). The levels of activated Cdr2 increase as cells grow, raising the possibility that Pom1 might inhibit Cdr2 activation and downstream signaling events in a size-dependent manner (Deng et al, 2014).…”

Section: Pom1 Suppresses Wee1 Node Bursts In Small Cellsmentioning

confidence: 99%

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Cell size-dependent regulation of Wee1 localization by Cdr2 cortical nodes

Allard

Opalko

Liu

et al. 2017

Preprint

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Cell size control requires mechanisms that link cell growth with Cdk1 activity. In fission yeast, the protein kinase Cdr2 forms cortical nodes that include the Cdk1 inhibitor Wee1, along with the Wee1-inhibitory kinase Cdr1. We investigated how nodes inhibit Wee1 during cell growth. Biochemical fractionation revealed that Cdr2 nodes were megadalton structures enriched for activated Cdr2, which increases in level during interphase growth. In live-cell TIRF movies, Cdr2 and Cdr1 remained constant at nodes over time, but Wee1 localized to nodes in short bursts. Recruitment of Wee1 to nodes required Cdr2 kinase activity and the noncatalytic N-terminus of Wee1. Bursts of Wee1 localization to nodes increased 20-fold as cells doubled in size throughout G2. Size-dependent signaling was due in part to the Cdr2 inhibitor Pom1, which suppressed Wee1 node bursts in small cells. Thus, increasing Cdr2 activity during cell growth promotes Wee1 localization to nodes, where inhibitory phosphorylation of Wee1 by Cdr1 and Cdr2 kinases promotes mitotic entry.SummaryCells turn off the mitotic inhibitor Wee1 to enter into mitosis. This study shows how cell growth progressively inhibits fission yeast Wee1 through dynamic bursts of localization to cortical node structures that contain Wee1 inhibitory kinases.

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Section: Wee1 Localization To Nodes Requires Cdr2 Kinase Activity Andsupporting

confidence: 53%

Section: Results and Discussion: Nodes Are Stable Complexes That Prommentioning

confidence: 71%

Section: Results and Discussion: Nodes Are Stable Complexes That Prommentioning

confidence: 99%

Section: Pom1 Suppresses Wee1 Node Bursts In Small Cellsmentioning

confidence: 99%

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Cell size-dependent regulation of Wee1 localization by Cdr2 cortical nodes

Allard

Opalko

Liu

et al. 2017

Preprint

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“…6D) (39), but our results suggest that a significant fraction of the protein is not phosphorylated in its activation loop and is therefore inactive. This suggests that all nodes are not necessarily catalytically active structures, consistent with some kinase-independent functions for Cdr2 (39,(53)(54)(55). Future studies should consider the dynamic Cdr2 activation system as a critical regulatory layer that operates in addition to node formation.…”

Section: Discussionmentioning

confidence: 83%

Phosphatases Generate Signal Specificity Downstream of Ssp1 Kinase in Fission Yeast

Deng

Lee

Schutt

et al. 2017

Molecular and Cellular Biology

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AMPK-related protein kinases (ARKs) coordinate cell growth, proliferation, and migration with environmental status. It is unclear how specific ARKs are activated at specific times. In the fission yeast Schizosaccharomyces pombe, the CaMKKlike protein kinase Ssp1 promotes cell cycle progression by activating the ARK Cdr2 according to cell growth signals. Here, we demonstrate that Ssp1 activates a second ARK, Ssp2/AMPK␣, for cell proliferation in low environmental glucose. Ssp1 activates these two related targets by the same biochemical mechanism: direct phosphorylation of a conserved residue in the activation loop (Cdr2-T166 and Ssp2-T189). Despite a shared upstream kinase and similar phosphorylation sites, Cdr2 and Ssp2 have distinct regulatory input cues and distinct functional outputs. We investigated this specificity and found that distinct protein phosphatases counteract Ssp1 activity toward its different substrates. We identified the PP6 family phosphatase Ppe1 as the primary phosphatase for Ssp2-T189 dephosphorylation. The phosphatase inhibitor Sds23 acts upstream of PP6 to regulate Ssp2-T189 phosphorylation in a manner that depends on energy but not on the intact AMPK heterotrimer. In contrast, Cdr2-T166 phosphorylation is regulated by protein phosphatase 2A but not by the Sds23-PP6 pathway. Thus, our study provides a phosphatase-driven mechanism to induce specific physiological responses downstream of a master protein kinase.KEYWORDS AMPK, Cdr2, Ssp1, fission yeast, kinase, phosphatase, pombe C ells control proliferation, polarized growth, and migration according to nutrient and environmental status. These processes must be tightly regulated during normal growth and development, and loss of this coordination is coupled with distinct steps in the initiation and maturation of tumors (1, 2). The AMP-activated protein kinase (AMPK)-related kinase (ARK) subfamily of protein kinases is essential for coordination of cell growth, proliferation, and migration with environmental status (3). Human cells express at least 13 members of this conserved kinase subfamily, including the metabolic sensor AMPK, the cytoskeletal kinase MARK1/Par-1, and the SAD kinases that regulate cell growth and polarity (4). Given their functions in diverse cell biological processes, it is remarkable that all ARKs can be activated by a shared upstream activating kinase, which phosphorylates the activation loop of ARK kinase domains to turn them "on" (4). To meet the needs of a rapidly changing environment, different ARKs need to be turned on and off at different times. This leads to a simple question: how does a common activation mechanism activate different ARKs at different times?The defining member of the ARK subfamily is AMPK. This conserved heterotrimeric complex is activated by low cellular energy status, typically the result of low levels of environmental nutrients (5). AMPK activation shifts cells toward catabolism and promotes the Warburg effect in cancer cells (5-7). The AMPK heterotrimer consists of a catalytic ␣ subun...

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Molecular mechanisms of contractile-ring constriction and membrane trafficking in cytokinesis

Gerien

2018

Biophys Rev

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In this review, we discuss the molecular mechanisms of cytokinesis from plants to humans, with a focus on contribution of membrane trafficking to cytokinesis. Selection of the division site in fungi, metazoans, and plants is reviewed, as well as the assembly and constriction of a contractile ring in fungi and metazoans. We also provide an introduction to exocytosis and endocytosis, and discuss how they contribute to successful cytokinesis in eukaryotic cells. The conservation in the coordination of membrane deposition and cytoskeleton during cytokinesis in fungi, metazoans, and plants is highlighted.

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Molecular control of the Wee1 regulatory pathway by the SAD kinase Cdr2

Cited by 17 publications

References 81 publications

Cell size-dependent regulation of Wee1 localization by Cdr2 cortical nodes

Cell size-dependent regulation of Wee1 localization by Cdr2 cortical nodes

Phosphatases Generate Signal Specificity Downstream of Ssp1 Kinase in Fission Yeast

Molecular mechanisms of contractile-ring constriction and membrane trafficking in cytokinesis

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