Downregulation of PP2ACdc55 Phosphatase by Separase Initiates Mitotic Exit in Budding Yeast

Queralt, Ethel; Lehane, Chris; Novák, Béla; Uhlmann, Frank

doi:10.1016/j.cell.2006.03.038

Cited by 233 publications

(395 citation statements)

References 53 publications

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“…In addition, we envision the approach to be beneficial in characterizing the cross-talk between phosphorylation and apoptotic proteolysis by including either phosphorylation mimetic or nonphosphorylatable residues in the TevS allele series (40,41). The approach outlined with the PTGR and SNIPer technology may prove particularly beneficial in profiling site-specific proteolysis throughout cell biology: for example, during cell-cycle progression mediated by separase during anaphase (42), differentiation, and intercellular signaling by Notch proteolysis (43), and mediating the unfolded protein response by ATF6 proteolysis (44).…”

Section: Discussionmentioning

confidence: 99%

Engineered cellular gene-replacement platform for selective and inducible proteolytic profiling

Morgan

Diaz

Zeitlin

et al. 2015

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

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Cellular demolition during apoptosis is completed by executioner caspases, that selectively cleave more than 1,500 proteins but whose individual roles are challenging to assess. Here, we used an optimized site-specific and inducible protease to examine the role of a classic apoptotic node, the caspase-activated DNase (CAD). CAD is activated when caspases cleave its endogenous inhibitor ICAD, resulting in the characteristic DNA laddering of apoptosis. We describe a posttranscriptional gene replacement (PTGR) approach where endogenous biallelic ICAD is knocked down and simultaneously replaced with an engineered allele that is susceptible to inducible cleavage by tobacco etch virus protease. Remarkably, selective activation of CAD alone does not induce cell death, although hallmarks of DNA damage are detected in human cancer cell lines. Our data strongly support that the highly cooperative action of CAD and inhibition of DNA repair systems are critical for the DNA laddering phenotype in apoptosis. Furthermore, the PTGR approach provides a general means for replacing wild-type protein function with a precisely engineered mutant at the transcriptional level that should be useful for cell engineering studies.DNA damage | apoptosis | ICAD | site-specific proteolysis |

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Section: Discussionmentioning

confidence: 99%

Engineered cellular gene-replacement platform for selective and inducible proteolytic profiling

Morgan

Diaz

Zeitlin

et al. 2015

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

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“…In budding yeast, PP2A has also been implicated in regulating mitotic exit and the spindle orientation checkpoint (Queralt et al 2006;Wang and Ng 2006;Chan and Amon 2009). Studies in other systems have reported that some forms of PP2A are inactivated to permit entry into mitosis and that these are important subsequently for mitotic exit (Barr et al 2011).…”

Section: Pp2a and Sin Signalingmentioning

confidence: 99%

Characterization of ypa1 and ypa2, the Schizosaccharomyces pombe Orthologs of the Peptidyl Proyl Isomerases That Activate PP2A, Reveals a Role for Ypa2p in the Regulation of Cytokinesis

Goyal

2012

Genetics

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The Schizosaccharomyces pombe septation initiation network (SIN) regulates cytokinesis. Cdc7p is the first kinase in the core SIN; we have screened genetically for SIN regulators by isolating cold-sensitive suppressors of cdc7-24. Our screen yielded a mutant in SPAC1782.05, one of the two fission yeast orthologs of mammalian phosphotyrosyl phosphatase activator. We have characterized this gene and its ortholog SPAC4F10.04, which we have named ypa2 and ypa1, respectively. We find that Ypa2p is the major form of protein phosphatase type 2A activator in S. pombe. A double ypa1-D ypa2-D null mutant is inviable, indicating that the two gene products have at least one essential overlapping function. Individually, the ypa1 and ypa2 genes are essential for survival only at low temperatures. The ypa2-D mutant divides at a reduced cell size and displays aberrant cell morphology and cytokinesis. Genetic analysis implicates Ypa2p as an inhibitor of the septation initiation network. We also isolated a cold-sensitive allele of ppa2, the major protein phosphatase type 2A catalytic subunit, implicating this enzyme as a regulator of the septation initiation network. SCHIZOSACCHAROMYCES pombe cells take the form of a cylinder capped by hemispherical ends. During interphase, cells grow mainly at their tips. Cell elongation ceases after mitotic commitment, and the cytoskeleton is reorganized in preparation for nuclear and cell division (reviewed by McCully and Robinow 1971;Mitchison and Nurse 1985;Marks et al. 1986;Hagan and Hyams 1988;Hagan 1998). The cell divides by fission after forming a septum in the middle of the cell. The site of division is defined at the onset of mitosis (Daga and Chang 2005), by the formation of an actomyosin contractile ring (CAR), whose assembly continues throughout mitosis (Wu et al. 2003;Pollard 2008;Pollard and Wu 2010). At the end of anaphase, the CAR is thought to guide synthesis of the multilayered division septum (reviewed by (Ishiguro 1998;Sipiczki 2007). Septation Initiation NetworkThe S. pombe septation initiation network (SIN) is a network of protein kinases that is essential for cytokinesis in the mitotic cycle; reviewed by (Gould and Simanis 1997;Simanis 2003;Wolfe and Gould 2005;Goyal et al. 2011). It is tightly regulated through the mitotic cell cycle to assure proper coordination of mitosis and cytokinesis, and during meiosis, where it is essential for spore formation (Krapp et al. 2006). Signaling requires the activity of three protein kinases, each of which has a regulatory subunit (kinase regulator); Cdc7p-Spg1p (Fankhauser and Simanis 1994;Schmidt et al. 1997;Mehta and Gould 2006), Sid1p-Cdc14p (Fankhauser and Simanis 1993;Guertin et al. 2000;Guertin and McCollum 2001), and Sid2p-Mob1p (Sparks et al. 1999;Hou et al. 2000;Salimova et al. 2000). SIN signaling is modulated by the nucleotide status of the GTPase Spg1p (Schmidt et al. 1997;Sohrmann et al. 1998), which is determined by the balance of spontaneous nucleotide exchange, a putative GEF, Etd1p Garcia-Cortes and McCollu...

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“…Because the FEAR controls Cdc14 release by inhibiting PP2A Cdc55 (9), the FEAR pathway is hyperactive in cdc55⌬ mutants, wherein the B-regulatory subunit of PP2A is deleted (7,8). To further confirm that CLB5 overexpression is toxic to FEAR mutants, we examined the growth of FEAR mutants overexpressing CLB5 in the absence of Cdc55.…”

Section: Fear Mutants Are More Sensitive To High Levels Of Clb5mentioning

confidence: 99%

Temporal control of the dephosphorylation of Cdk substrates by mitotic exit pathways in budding yeast

Jin

Liu²,

Liang³

et al. 2008

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

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The temporal phosphorylation of cell cycle-related proteins by cyclin-dependent kinases (Cdks) is critical for the correct order of cell cycle events. In budding yeast, CDC28 encodes the only Cdk and its association with various cyclins governs the temporal phosphorylation of Cdk substrates. S-phase Cdk substrates are phosphorylated earlier than mitotic Cdk substrates, which ensures the sequential order of DNA synthesis and mitosis. However, it remains unclear whether Cdk substrates are dephosphorylated in temporally distinct windows. Cdc14 is a conserved protein phosphatase responsible for the dephosphorylation of Cdk substrates. In budding yeast, FEAR (Cdc14 early anaphase release) and MEN (mitotic exit network) activate phosphatase Cdc14 by promoting its release from the nucleolus in early and late anaphase, respectively. Here, we show that the sequential Cdc14 release and the distinct degradation timing of different cyclins provides the molecular basis for the differential dephosphorylation windows of S-phase and mitotic cyclin substrates. Our data also indicate that FEAR-induced dephosphorylation of S-phase Cdk substrates facilitates anaphase progression, revealing an extra layer of mitotic regulation.Cdc14 phosphatase ͉ spindle ͉ Clb5 ͉ Clb2 ͉ anaphase

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Downregulation of PP2ACdc55 Phosphatase by Separase Initiates Mitotic Exit in Budding Yeast

Cited by 233 publications

References 53 publications

Engineered cellular gene-replacement platform for selective and inducible proteolytic profiling

Engineered cellular gene-replacement platform for selective and inducible proteolytic profiling

Characterization of ypa1 and ypa2, the Schizosaccharomyces pombe Orthologs of the Peptidyl Proyl Isomerases That Activate PP2A, Reveals a Role for Ypa2p in the Regulation of Cytokinesis

Temporal control of the dephosphorylation of Cdk substrates by mitotic exit pathways in budding yeast

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