Crm1-Mediated Nuclear Export of Cdc14 is Required for the Completion of Cytokinesis in Budding Yeast

Bembenek, Joshua N.; Kang, Jungseog; Kurischko, Cornelia; Li, Bing; Raab, Jesse R.; Belanger, Kenneth D.; Luca, Francis C.; Yu, Hongtao

doi:10.4161/cc.4.7.1798

Cited by 48 publications

(67 citation statements)

References 65 publications

(99 reference statements)

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“…Mutations within the NES of Cdc14 were reported to allow destruction of mitotic cyclins but block cytokinesis (23). Using this conditional, separation-of-function mutant, we investigated the effect of Cdc14 release when it is incapable of reaching cytoplasmic targets.…”

Section: Journal Of Biological Chemistrymentioning

confidence: 99%

“…In cells where Cdk1 activity is ectopically inactivated without Cdc14 release from the nucleolus, the actomyosin ring shows defects in contraction and cell separation (21,22). Interestingly, Cdc14 localizes to the bud neck in late mitosis, and cells carrying Cdc14 with a mutated nuclear export sequence (NES) fail to localize Cdc14 to the bud neck or contract the actomyosin ring at restrictive temperatures (23). Thus, it is likely that Cdc14 has cytoplasmic targets at the bud neck whose dephosphorylation is essential for cytokinesis.…”

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confidence: 99%

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Global Analysis of Cdc14 Phosphatase Reveals Diverse Roles in Mitotic Processes

Bloom

Cristea

Procko

et al. 2011

Journal of Biological Chemistry

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Cdc14 phosphatase regulates multiple events during anaphase and is essential for mitotic exit in budding yeast. Cdc14 is regulated in both a spatial and temporal manner. It is sequestered in the nucleolus for most of the cell cycle by the nucleolar protein Net1 and is released into the nucleus and cytoplasm during anaphase. To identify novel binding partners of Cdc14, we used affinity purification of Cdc14 and mass spectrometric analysis of interacting proteins from strains in which Cdc14 localization or catalytic activity was altered. To alter Cdc14 localization, we used a strain deleted for NET1, which causes full release of Cdc14 from the nucleolus. To alter Cdc14 activity, we generated mutations in the active site of Cdc14 (C283S or D253A), which allow binding of substrates, but not dephosphorylation, by Cdc14. Using this strategy, we identified new interactors of Cdc14, including multiple proteins involved in mitotic events. A subset of these proteins displayed increased affinity for catalytically inactive mutants of Cdc14 compared with the wild-type version, suggesting they are likely substrates of Cdc14. We have also shown that several of the novel Cdc14-interacting proteins, including Kar9 (a protein that orients the mitotic spindle) and Bni1 and Bnr1 (formins that nucleate actin cables and may be important for actomyosin ring contraction) are specifically dephosphorylated by Cdc14 in vitro and in vivo. Our 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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Section: Journal Of Biological Chemistrymentioning

confidence: 99%

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confidence: 99%

Global Analysis of Cdc14 Phosphatase Reveals Diverse Roles in Mitotic Processes

Bloom

Cristea

Procko

et al. 2011

Journal of Biological Chemistry

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“…The second phase of Cdc14p release occurs during telophase and is dependent on other MEN proteins, such as Cdc5p, Tem1p, Cdc15p, and Dbf2p (Shou et al., 1999;Visintin et al, 1999). Once released in telophase, Cdc14p disperses throughout the cell and concentrates on spindle pole bodies (SPBs) and the bud neck ring (the site of cytokinesis) before returning to the nucleolus (Yoshida et al, 2002;Pereira and Schiebel, 2003;Bembenek et al, 2005 mitotic substrates for Cdc14p include Cdh1p, Sic1p, and Swi5p, which are CDK substrates that regulate mitotic cyclin degradation, CDK inactivation, and G1 gene expression, respectively (Visintin et al, 1998). Cdc14p was shown to regulate the kinetochore-associated chromosomal passenger proteins Sli15p, and Ipl1p kinase, which are homologues to mammalian INCENP and Aurora B kinase (Pereira and Schiebel, 2003;Vagnarelli and Earnshaw, 2004).…”

Section: Introductionmentioning

confidence: 99%

The Mitotic Exit Network Mob1p-Dbf2p Kinase Complex Localizes to the Nucleus and Regulates Passenger Protein Localization

Stoepel

Ottey

Kurischko

et al. 2005

MBoC

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The Saccharomyces cerevisiae mitotic exit network (MEN) is a conserved signaling network that coordinates CDK inactivation, cytokinesis and G1 gene transcription. The MEN Cdc14p phosphatase is sequestered in the nucleolus and transiently released in early anaphase and telophase. Cdc14p mediates mitotic exit by dephosphorylating Cdk1p substrates and promoting Cdk1p inactivation. Cdc14p also regulates the localization of chromosomal passenger proteins, which redistribute from kinetochores to the mitotic spindle during anaphase. Here we present evidence that the MEN protein kinase complex Mob1p-Dbf2p localizes to mitotic nuclei and partially colocalizes with Cdc14p and kinetochore proteins. Chromatin immunoprecipitation (ChIP) experiments reveal that Mob1p, Dbf2p, and Cdc14p associate with centromere DNA and require the centromere binding protein Ndc10p for this association. We establish that Mob1p is essential for maintaining the localization of Aurora, INCENP, and Survivin chromosomal passenger proteins on anaphase spindles, whereas Cdc14p and the Mob1p-Dbf2p-activating kinase Cdc15p are required for establishing passenger protein localization on the spindle. Moreover, Mob1p, but not Cdc15p, is required for dissociating Aurora from the kinetochore region. These findings reveal kinetochores as sites for MEN signaling and implicate MEN in coordinating chromosome segregation and/or spindle integrity with mitotic exit and cytokinesis via regulation of chromosome passenger proteins. INTRODUCTIONThe Saccharomyces cerevisiae mitotic exit network (MEN) is a conserved signaling network that coordinates several events associated with mitotic exit, including the inactivation of cyclin dependent kinase (CDK), cytokinesis activation, initiation of G1 gene transcription, and formation of prereplication complexes (Seshan and Amon, 2004;Simanis, 2003). MEN is comprised of several regulatory proteins, including Tem1p GTPase, a two-component GTPase-activating factor (GAP) Bub1p-Bfa1p, a putative GTP exchange factor (GEF) Lte1p, Cdc14p protein phosphatase, four protein kinases Cdc5p, Cdc15p, Dbf2p, Dbf20p, and a Dbf2p-associated protein Mob1p. The equivalent signaling network in Schizosaccharomyces pombe is called the septation initiation network (SIN) and is required for coordinating cytokinesis with CDK inactivation (Simanis, 2003;Wolfe and Gould, 2004). Most MEN/SIN proteins are conserved from yeast to human, however the functions of MEN/SIN-related networks have not been resolved in other eukaryotes.S. cerevisiae Cdc14p plays a key role in MEN signaling (reviewed in Seshan and Amon, 2004;Stegmeier and Amon, 2004). Cdc14p is a temporally and spatially regulated proline directed phosphatase that directly mediates mitotic exit by promoting CDK inactivation and dephosphorylating CDK substrates (Gray et al., 2003;Visintin et al., 1998). Cdc14p is sequestered in the nucleolus throughout most of the cell cycle by binding to the nucleolar RENT complex (Shou et al., 1999;Visintin et al., 1999). Its release from the nucleolus is essentia...

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“…Dbf2/Mob1 phosphorylates Cdc14 to inactivate its nuclear localization signal (NLS), promoting release of Cdc14 into the cytoplasm (46). Cytoplasmic Cdc14 dephosphorylates CDK targets to promote mitotic exit and the final events of cell division (7,61,67).…”

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confidence: 99%

Mitotic Exit Control of the Saccharomyces cerevisiae Ndr/LATS Kinase Cbk1 Regulates Daughter Cell Separation after Cytokinesis

Brace

Hsu

Weiss

2011

Molecular and Cellular Biology

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Saccharomyces cerevisiae cell division ends with destruction of a septum deposited during cytokinesis; this must occur only after the structure's construction is complete. Genes involved in septum destruction are induced by the transcription factor Ace2, which is activated by the kinase Cbk1, an Ndr/LATS-related protein that functions in a system related to metazoan hippo pathways. Phosphorylation of a conserved hydrophobic motif (HM) site regulates Cbk1; at peak levels in late mitosis we found that approximately 3% of Cbk1 carries this modification. HM site phosphorylation prior to mitotic exit occurs in response to activation of the FEAR (Cdc fourteen early anaphase release) pathway. However, HM site phosphorylation is not sufficient for Cbk1 to act on Ace2: the kinase is also negatively regulated prior to cytokinesis, likely by cyclin-dependent kinase (CDK) phosphorylation. Cbk1 cannot phosphorylate Ace2 until after mitotic exit network (MEN)-initiated release of the phosphatase Cdc14. Treatment of Cbk1 with Cdc14 in vitro does not increase its intrinsic enzymatic activity, but Cdc14 is required for Cbk1 function in vivo. Thus, we propose that Cdc14 coordinates cell separation with mitotic exit via FEAR-initiated phosphorylation of the Cbk1 HM site and MEN-activated reversal of mitotic CDK phosphorylations that block both Cbk1 and Ace2 function.

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Crm1-Mediated Nuclear Export of Cdc14 is Required for the Completion of Cytokinesis in Budding Yeast

Cited by 48 publications

References 65 publications

Global Analysis of Cdc14 Phosphatase Reveals Diverse Roles in Mitotic Processes

Global Analysis of Cdc14 Phosphatase Reveals Diverse Roles in Mitotic Processes

The Mitotic Exit Network Mob1p-Dbf2p Kinase Complex Localizes to the Nucleus and Regulates Passenger Protein Localization

Mitotic Exit Control of the Saccharomyces cerevisiae Ndr/LATS Kinase Cbk1 Regulates Daughter Cell Separation after Cytokinesis

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