Phosphorylation by Cyclin B-Cdk Underlies Release of Mitotic Exit Activator Cdc14 from the Nucleolus

Azzam, Ramzi; Chen, Susan L.; Shou, Wenying; Mah, Angie S.; Alexandru, Gabriela; Nasmyth, Kim; Annan, Roland S.; Carr, Steven A.; Deshaies, Raymond J.

doi:10.1126/science.1099402

Cited by 171 publications

(251 citation statements)

References 21 publications

Supporting

Mentioning

233

Contrasting

Unclassified

Order By: Relevance

“…(B) Morphology of Dictyostelium cells. The phase-contrast image is shown in a, c, and e; b, d, and f show nuclear nucleolus (28). It is possible that cdc14 dephosphorylates MKLP1, causing it to bind to microtubules, which results in localization of mitotic machinery (including INCENP-Aurora B) to the spindle (29).…”

Section: Discussionmentioning

confidence: 99%

A mitotic kinesin-like protein required for normal karyokinesis, myosin localization to the furrow, and cytokinesis in Dictyostelium

Lakshmikanth

Warrick²,

Spudich³

2004

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Dictyostelium mitotic kinesin Kif12 is required for cytokinesis. Myosin II localization to the cleavage furrow is severely depressed in Kif12-null (⌬kif12) cells, which accounts in part for the cytokinesis failure. Myosin II-null cells, however, undergo mitosis-coupled cytokinesis when adhering to a surface, whereas the ⌬kif12 cells cannot. During mitosis, the rate of change of internuclear separation in ⌬kif12 cells is reduced compared with wild-type cells, indicating multiple roles of this molecular motor during mitosis and cytokinesis. GFP-Kif12, which rescues wild-type behavior when expressed in the ⌬kif12 strain, is concentrated in the nucleus in interphase cells, translocates to the cytoplasm at the onset of mitosis, appears in the centrosomes and spindle, and later is concentrated in the spindle midbody. Given these results, we hypothesize a mechanism for myosin II translocation to the furrow to set up the contractile ring.Kif12 ͉ cell-cycle regulation ͉ mitosis C ytokinesis and karyokinesis are fundamental to cell division and propagation. Karyokinesis involves nuclear segregation and requires the formation of a microtubule network on which the chromosomes are segregated with the help of kinesin-based motors. Cytokinesis involves division of cellular components of a parental cell into two daughter cells, and it is precisely timed along with nuclear division in almost all cells. Cytokinesis is generally achieved by an actin-myosin-based contractile ring that is assembled along the plane perpendicular to the axis of the spindle poles. Karyokinesis and cytokinesis are tightly synchronized to ensure high fidelity of genomic information transfer.Classic experiments carried out on sand dollar eggs established the importance of the mitotic spindles and astral microtubules in setting up the contractile ring (1, 2). The central spindle has been shown to be important for the completion of cytokinesis in several organisms, including Dictyostelium (3). Although a number of proteins have been shown to be associated with the mitotic spindle and implicated in cytokinesis (4-6), the molecular clues for the crosstalk between the spindle and cortical midzone during mitosis remain elusive.Members of the mitotic kinesin family have been shown to play key roles in the assembly and function of the mitotic spindle. In mammalian cells, MKLP1 is required for the formation of the midbody matrix and the completion of cytokinesis (7,8). Similar gene disruption experiments of MKLP1 orthologs in zebrafish (9), Drosophila (10), and Caenorhabditis elegans (11) also result in a cytokinesis failure. In most cases, furrow ingression is initiated, but cytokinesis fails to complete. Thus, MKLP1 is a key player in passing information from the mitotic spindle to the cleavage furrow.Myosin II is a major component of the cellular machinery responsible for cytokinesis (12, 13). However, little is known about how myosin is translocated to the cleavage furrow during mitosis. Using Dictyostelium as a model system, our laboratory has been studying m...

show abstract

Section: Discussionmentioning

confidence: 99%

A mitotic kinesin-like protein required for normal karyokinesis, myosin localization to the furrow, and cytokinesis in Dictyostelium

Lakshmikanth

Warrick²,

Spudich³

2004

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…C283S or Cdc14-D253A appeared to be slightly increased compared with wild-type Cdc14. This may reflect that Net1 is an in vitro substrate of Cdc14, although hypophosphorylated Net1 has been shown to have an increased affinity for Cdc14 (44,50).…”

Section: Figure 2 a Subset Of Cdc14 Interactors Display Enhanced Binmentioning

confidence: 99%

Global Analysis of Cdc14 Phosphatase Reveals Diverse Roles in Mitotic Processes

Bloom

Cristea

Procko

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

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.

show abstract

“…Furthermore, mutations in Cdc5p phosphorylation sites on Net1p weaken Net1p-Cdc14p association (Shou et al, 2002). Recent data suggest that FEAR proteins induce Cdc14p release by promoting Cdk1p phosphorylation of Net1p/Cfi1p (Azzam et al, 2004). The MEN-dependent mechanism for Cdc14p release during telophase remains a mystery.…”

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

View full text Add to dashboard Cite

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...

show abstract

Phosphorylation by Cyclin B-Cdk Underlies Release of Mitotic Exit Activator Cdc14 from the Nucleolus

Cited by 171 publications

References 21 publications

A mitotic kinesin-like protein required for normal karyokinesis, myosin localization to the furrow, and cytokinesis in Dictyostelium

A mitotic kinesin-like protein required for normal karyokinesis, myosin localization to the furrow, and cytokinesis in Dictyostelium

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

Contact Info

Product

Resources

About