APC/C-Cdh1-dependent anaphase and telophase progression during mitotic slippage
© Toda et al; licensee BioMed Central Ltd. 2012
Received: 1 September 2011
Accepted: 9 February 2012
Published: 9 February 2012
The spindle assembly checkpoint (SAC) inhibits anaphase progression in the presence of insufficient kinetochore-microtubule attachments, but cells can eventually override mitotic arrest by a process known as mitotic slippage or adaptation. This is a problem for cancer chemotherapy using microtubule poisons.
Here we describe mitotic slippage in yeast bub2Δ mutant cells that are defective in the repression of precocious telophase onset (mitotic exit). Precocious activation of anaphase promoting complex/cyclosome (APC/C)-Cdh1 caused mitotic slippage in the presence of nocodazole, while the SAC was still active. APC/C-Cdh1, but not APC/C-Cdc20, triggered anaphase progression (securin degradation, separase-mediated cohesin cleavage, sister-chromatid separation and chromosome missegregation), in addition to telophase onset (mitotic exit), during mitotic slippage. This demonstrates that an inhibitory system not only of APC/C-Cdc20 but also of APC/C-Cdh1 is critical for accurate chromosome segregation in the presence of insufficient kinetochore-microtubule attachments.
The sequential activation of APC/C-Cdc20 to APC/C-Cdh1 during mitosis is central to accurate mitosis. Precocious activation of APC/C-Cdh1 in metaphase (pre-anaphase) causes mitotic slippage in SAC-activated cells. For the prevention of mitotic slippage, concomitant inhibition of APC/C-Cdh1 may be effective for tumor therapy with mitotic spindle poisons in humans.
KeywordsAnaphase promoting complex/cyclosome (APC/C) Bub2 Cdh1 mitotic exit network (MEN) mitotic slippage Saccharomyces cerevisiae securin
The anaphase-promoting complex/cyclosome (APC/C) is an E3 ubiquitin ligase that plays a major role in cell cycle control by targeting substrates for proteasomal degradation. The complex is activated by two WD40 activator proteins, Cdc20/Fizzy/Fzy or Cdh1/Fizzy-related/Fzr. This destruction is strictly ordered to ensure that cell cycle events are executed in a timely fashion [1–5]. Whereas APC/C-Cdc20 is activated at metaphase-anaphase transition, APC/C-Cdh1 is activated after APC/C-Cdc20 activation. In the budding yeast Saccharomyces cerevisiae, APC/C-Cdh1 is activated from telophase to late G1 phase [6, 7]. The switch from APC/C-Cdc20 to APC/C-Cdh1 is regulated by multiple mechanisms [5, 8–10]: Cyclin B-Cdk1 (cyclin-dependent kinase) inhibits Cdh1 activation in metaphase, but cyclin B degradation mediated by APC/C in late M phase reduces cyclin B-Cdk1 activity, leading to Cdh1 activation. In addition, APC/C-Cdh1 mediates Cdc20 degradation, thereby promoting switching from APC/C-Cdc20 to APC/C-Cdh1.
The spindle assembly checkpoint (SAC) ensures faithful chromosome segregation during cell division [11, 12]. In the presence of insufficient kinetochore-microtubule attachments, the SAC inhibits anaphase onset by the inhibition of APC/C-Cdc20. The SAC recruits checkpoint proteins, including Mad1, Mad2, Bub1, BubR1 (Mad3 in yeast), Bub3 and Mps1, to unattached kinetochores. As a result, Mad2, BubR1 and Bub3 bind to and suppress APC/C-Cdc20 and form the mitotic checkpoint complex (MCC) . Once all chromosomes have achieved proper kinetochore-microtubule attachment, checkpoint signaling ceases, which is called SAC deactivation or inactivation, and Mad2/BubR1/Bub3 are released from APC/C-Cdc20. It allows active APC/C-Cdc20 to drive cells into anaphase by inducing the degradation of securin and cyclin B. The degradation of securin permits sister-chromatid separation, and the destruction of cyclin B reduces Cdk1 activity. In contrast to the molecular mechanisms of the SAC activation, those of SAC deactivation are poorly understood [14, 15].
Microtubule targeted drugs are of clinical importance in the successful treatment of a variety of human cancers because they activate the SAC and induce mitotic arrest that leads to apoptotic cell death . However, in the continued presence of conditions that normally keep the SAC active, some cells escape from mitosis, resulting in tetraploid cells [16, 17]. This phenomenon is termed mitotic slippage or adaptation. This process is largely responsible for the failure to efficiently block tumor progression. Mitotic slippage depends on progressive degradation of cyclin B, while the SAC is active, indicating that mitotic slippage occurs through the overriding of activated SAC signaling [18, 19]. Mitotic exit occurs once cyclin B-Cdk1 activity has decreased below a critical threshold required to maintain a mitotic state . In addition to cyclin B, other mitotic APC/C substrates, including securin, are also degraded during mitotic slippage, and a double knockdown of Cdc20 and Cdh1 prevents the degradation of APC/C substrates during mitotic slippage . These findings indicate that APC/C is critical for mitotic slippage. However, which protein does mitotic slippage require, Cdc20 or Cdh1? Furthermore, how can APC/C be activated, although the SAC is active? The degradation of Cyclin A and NIMA-related kinase 2A (Nek2A) in early mitosis is dependent on APC/C-Cdc20, and this process is not inhibited by the SAC . While the SAC-dependent substrate cyclin B requires Cdc20 for recruitment to APC/C, Nek2A can bind the APC/C in the absence of Cdc20 . Thus, the SAC suppresses the degradation of most, but not all, substrates of APC/C-Cdc20. However, degradation of cyclin A and Nek2A does not trigger metaphase-anaphase transition and mitotic slippage. It is unclear how mitotic exit (telophase onset) can be initiated in metaphase-arrested cells during mitotic slippage; less attention has been paid to how anaphase is executed during mitotic slippage.
In budding yeast, mitotic slippage-like phenomena have been reported, but they are relatively ill-defined, as compared with mammalian cells, because the SAC status is obscure. It is important to determine the SAC status during mitotic slippage (and slippage-like phenomena), in order to distinguish mitotic slippage that overrides the activated SAC from events caused by SAC deactivation. Mitotic exit accompanied by securin degradation, sister-chromatid separation and nuclear division was found after treatment of the wild-type yeast cells with the microtubule depolymerizer benomyl but not with nocodazole . It is unknown whether these phenomena found in the presence of benomyl are indeed mitotic slippage, because the SAC status has not been characterized. Interestingly, mitotic slippage (or slippage-like phenomena) is prominently observed in mutant cells deficient in the budding uninhibited by benzimidazole (BUB) 2 gene in the presence of nocodazole. Among BUB proteins, whereas Bub1 and Bub3 are components of the SAC, Bub2 is an inhibitor of the mitotic exit network (MEN) that promotes anaphase-telophase transition [8–10, 24, 25]. Although bub2Δ cells exhibit an intact SAC, they fail to arrest in metaphase and exit from mitosis in the presence of nocodazole, leading to cell death [24, 26–28]. In addition, bub2Δ cells cannot effectively arrest at metaphase when the SAC is activated by MAD2 overexpression .
Nocodazole-treated bub2Δ cells exhibit securin degradation, sister-chromatid separation (indexes of anaphase progression) and rebudding (an index of telophase progression and mitotic exit) [24, 26, 27]. Thus, anaphase and telophase progression occurs in nocodazole-treated bub2Δ cells. A mutation in the MEN factor Tem1 suppresses bub2Δ-induced sister-chromatid segregation . It suggests that precocious activation of the MEN causes mitotic slippage, but the molecular mechanism responsible is largely unknown. Although securin degradation and sister-chromatid separation are normally mediated by APC/C-Cdc20 at anaphase onset, these events found in nocodazole-treated bub2Δ cells are not repressed by a cdc20-3 mutation at a restrictive temperature . These findings indicate that anaphase progression in nocodazole-treated bub2Δ cells occurs independently of APC/C-Cdc20. On the other hand, sister-chromatid separation in nocodazole-bub2Δ cells is suppressed by a lack of the APC/C core subunit Cdc26 , suggesting that APC/C itself is required for bub2Δ-mediated anaphase progression. Thus, the bub2Δ stain might be a useful model for mitotic slippage. We show herein that nocodazole-treated bub2Δ cells override the active SAC-mediated metaphase arrest and cause securin degradation, sister-chromatid separation and mitotic exit and that APC/C-Cdh1 is critical for mitotic slippage.
Precocious activation of the MEN induces mitotic slippage
Given the role of Bub2, it is most likely that precocious MEN activation occurs in bub2Δ cells, leading to mitotic slippage. To assess this idea, we examined whether overexpression of the MEN inhibitor Bfa1 cancels mitotic slippage in bub2Δ cells. This was indeed the case: sister-chromatid separation and rebudding was completely repressed by BFA1 overexpression (Figures 1A-C, bub2Δ GAL-BFA1). This demonstrated that MEN activation causes anaphase and telophase onset during mitotic slippage. It was also reported that a mutation in the MEN factor Tem1 suppresses bub2Δ-induced sister-chromatid segregation .
Because nuclear division (an index of anaphase progression) is dependent on spindle microtubules, it is inhibited when microtubules are completely abrogated. However, nuclear division in the bub2Δ cells treated with nocodazole (LKT Laboratories, Lot No. QJ1275) was identified, although no detectable microtubules were found in the indirect immunoflorescence assay (data not shown). This finding suggested that there were imperceptible microtubules causing nuclear division. However, it was noteworthy that the SAC was still active under these conditions (see below). These findings indicated that nocodazole continues to activate the SAC sufficiently and that the phenomena found in bub2Δ cells here are caused by mitotic slippage but not SAC deactivation/inactivation.
We also examined mitotic slippage of bub2Δ cells when the SAC gene MAD2 was overexpressed. MAD2 overexpression causes SAC activation-mediated metaphase arrest, but during a long-term treatment cells override metaphase arrest and cause cell proliferation, although profiles of sister-chromatid separation and nuclear division, chromosome missegregation during mitotic slippage were not described . When MAD2 was overexpressed for 6 h, rebudding (mitotic exit) was frequently found in bub2Δ cells, as compared with wild-type cells (Additional file 1), which was consistent with the finding that cell proliferation was promoted by the bub2Δ mutation . Furthermore, it was found that sister-chromosome segregation and nuclear division were also prominent in bub2Δ cells (Additional file 1). Thus, both nocodazole treatment and MAD2 overexpression similarly caused mitotic slippage in bub2Δ cells. In contrast, chromosome missegregation in MAD2-overexpressing bub2Δ cells was not detectable (data not shown), which was probably because in this case microtubules were intact and a proper kinetochore-microtubule attachment was established.
APC/C-Cdh1 is critical for chromosome separation during mitotic slippage
We suspected that precocious MEN activation in nocodazole-treated bub2Δ cells causes activation of APC/C-Cdh1, leading to mitotic slippage. Indeed, a lack of Cdh1 markedly repressed sister-chromatid separation and nuclear division in nocodazole-treated bub2Δ cells (Figure 2A, B). These observations clearly indicated that APC/C-Cdh1, but not APC/C-Cdc20, is responsible for MEN-mediated anaphase progression. In contrast, rebudding (mitotic exit) was not suppressed by CDH1 deletion, probably because the CDK inhibitor Sic1 induced by the MEN also contributes to the repression of CDK activity and is sufficient for mitotic exit in cdh1Δ cells, as in normal mitosis.
APC/C-Cdh1-mediated securin degradation is required for sister-chromatid separation during mitotic slippage
Next, we examined whether this securin degradation during mitotic slippage is required for sister-chromatid separation. APC/C-Cdh1 targets securin through D- and KEN boxes [33, 34]. To test this, we ectopically expressed a non-degradable securin mutant devoid of both D- and KEN boxes (securin-dkb) . Securin-dkb strongly repressed sister-chromatid separation (Figures 3B, C). In contrast, as expected, expression of a securin mutant lacking only the D-box (securin-db) repressed sister-chromatid separation less effectively. These findings demonstrated that APC/C-Cdh1-mediated securin degradation is a prerequisite for sister-chromatid separation during mitotic slippage in nocodazole-treated bub2Δ cells.
Separase executes sister-chromatid separation and nucleolar segregation during mitotic slippage
In normal early anaphase, the liberated separase also causes nucleolar segregation in a manner independent of its protease activity [41–43]. The nucleolar segregation into mother and daughter cells was observed, together with nuclear division during mitotic slippage in the bub2Δ cells (Figure 4C). This indicates that protease-independent action of separase is also promoted during mitotic slippage.
The SAC is active during mitotic slippage
Ectopic activation of Cdh1 causes mitotic slippage
Cdc14 phosphatase, which antagonizes CDK, promotes APC/C-Cdh1 activation and mitotic exit in telophase [5, 9, 41]. CDC14 overexpression from G1 phase induced securin degradation (Figure 6C) but inhibited G1/S progression  (data not shown). Hence, we overexpressed CDC14 in metaphase-arrested cells treated with nocodazole. CDC14 overexpression promoted securin degradation and sister-chromatid separation (Figures 6D-F). However, both of these events were repressed in cdh1Δ cells (Figure 6E, F). These findings indicated that ectopic activation of Cdc14 causes mitotic slippage via APC/C-Cdh1. Consistent with the previous report that CDC14 overexpression promotes mitotic exit but represses budding in the next S phase, because of the counteraction of Cdc14 against CDK-mediated phosphorylation , no promotion of rebudding in the next S phase by CDC14 overexpression was observed (Figure 6F). Overall, these findings in Cdh1- and Cdc14-overexpressing cells supported the notion that precocious activation of APC/C-Cdh1 in pre-anaphase triggers mitotic slippage.
Precocious activation of APC/C-Cdh1 in metaphase causes mitotic slippage
APC/C-Cdh1-mediated anaphase progression during mitotic slippage had two prominent features. First, APC/C-Cdh1-mediated anaphase progression brought about chromosome missegregation, because APC/C-Cdh1 is not inhibited by the SAC in the presence of inappropriate kinetochore-microtubule attachments; therefore, APC/C-Cdh1-mediated securin degradation results in chromosome missegregation. This demonstrated that an inhibitory system not only of APC/C-Cdc20 but also of APC/C-Cdh1 is critical for accurate chromosome segregation in the presence of insufficient kinetochore-microtubule attachments.
Second, APC/C-Cdh1 simultaneously starts anaphase and telophase from metaphase. APC/C-Cdc20 recognizes the D-box of a relatively limited umber of targets (the important targets are only cyclin Clb5 and securin Pds1 in budding yeast) , whereas APC/C-Cdh1 recognizes various motifs on numerous targets (A-, O-, CRY and GxEN boxes, in addition to D- and KEN boxes) [2, 46–48]. Namely, APC/C-Cdh1 could target substrates for APC/C-Cdc20, which allowed simultaneous onsets of anaphase and telophase. APC/C-Cdh1 targets securin Pds1 in a manner dependent on D- and KEN boxes in vitro [33, 34], and ectopically expressed Pds1 in G1 phase was degraded in a manner dependent on APC/C-Cdh1 . These findings suggested that APC/C-Cdh1 mediates securin degradation from telophase to G1 phase in vivo. If APC/C-Cdh1 becomes activated abnormally in metaphase, it can target securin, leading to sister-chromatid separation (Figure 7B). This study clarifies these abnormal aspects of precocious APC/C-Cdh1 activation in metaphase cells and emphasizes that sequential activation of APC/C-Cdc20-to-APC/C-Cdh1 is critical for mitosis.
Deregulation of APC/C-Cdh1 in other cell phases brings about different outputs. CDH1 overexpression in asynchronized cells leads to elongated buds, G2 phase arrest, and 4C DNA content in some cells [6, 7, 50]. Precocious activation of APC/C-Cdh1 in G2 phase targets proteins that are required for separation of the spindle pole body (SPB, yeast centrosome), the BimC family kinesins Cin8/Eg5 and Kip1 and the interpolar microtubule midzone protein Ase1 [51, 52]. Thus, deregulation of Cdh1 activity compromises genome transmission in various ways and timely activation and inactivation of APC/C-Cdh1 are pivotal for accurate genome transmission.
APC/C-Cdh1-mediated mitotic slippage in other organisms
In fission yeast, the septation initiation network (SIN), a signaling pathway homologous to the MEN, coordinates mitosis and cytokinesis [8, 25, 53, 54]. Cdc16 (Bub2 ortholog) acts as a negative factor of the SIN, and cdc16 mutant cells undergo cytokinesis in the presence of the microtubule destabilizer thiabendazole [55–57]. This suggests that SIN-mediated APC/C-Cdh1/Ste9 activation causes mitotic slippage. In addition, the fission yeast securin Cut2 also has D- and KEN boxes (Figure 7C). We postulate that APC/C-Cdh1/Ste9-mediated securin degradation and sister-chromatid separation is promoted during mitotic slippage in fission yeast.
In mammalian mitosis, APC/C-Cdc20 and APC/C-Cdh1 are sequentially activated [3, 5, 58]. Mitotic slippage depends on progressive degradation of cyclin B with the SAC active [18, 19]. This suggests that APC/C-Cdh1, but not APC/C-Cdc20, is also involved in mitotic slippage in mammalian cells. Cdc20 and Cdh1 target securin in a manner dependent on D/KEN-boxes [59, 60] (see Figure 7C). This suggests that precocious activation of APC/C-Cdh1 similarly causes securin degradation and sister-chromatid separation during mitotic slippage in mammalian cells. In fact, deregulation of Cdh1 in pre-anaphase results in premature securin degradation and sister-chromatid separation [59, 61]. The present study predicts that for prevention of mitotic slippage, concomitant inhibition of APC/C-Cdh1 may be effective for tumor therapy with mitotic spindle poisons in humans.
The sequential activation of APC/C-Cdc20-to-APC/C-Cdh1 during mitosis is critical for accurate mitosis. Precocious activation of APC/C-Cdh1 in metaphase (pre-anaphase) causes mitotic slippage in microtubule poison-treated cells. For prevention of mitotic slippage, concomitant inhibition of APC/C-Cdh1 may be effective for tumor therapy with mitotic spindle poisons in human.
Strains, plasmids, media and materials
Yeast strains used in this study
Mata ura3 his3 leu2 trp1 ade2 can1 (lab stock)
SCU893 bub2::hphMX4 (this study)
SCU893 his3::GFP12-LacI12-NLS::HIS3 trp1::LacOx256-TRP1 (this study)
SCU397 (bub2Δ CEN-GFP)
SCU396 bub2::loxP (this study)
SCU893 ura3::tetO2x112::URA3 leu2::tetR-GFP-NLS::LEU2 (this study)
SCU399 (bub2Δ CEN-GFP)
SCU398 bub2::hphMX (this study)
SCU404 (bub2Δ GAL-SCC1-RRDD CEN-GFP)
SCU397 leu2::GAL1-SCC1-R180D/R268D-HA3::LEU2 (this study)
SCU408 (bub2Δ PDS1-HA3)
SCU151 PDS1-HA3::URA3 (this study)
SCU410 (bub2Δ MET3-CDC20 CEN-GFP)
SCU399 MET3-CDC20::TRP1 (this study)
SCU15 bar1::hisG (U. Surana)
SCU1226 (cdh1Δ CEN-GFP)
SCU15 ura3::tetO::URA3 leu2::tetR::LEU2 cdh1::HIS3 
SCU15 cdh1::kanR 
SCU1336 (bub2Δ cdh1Δ CEN-GFP)
SCU1226 bub2::kanMX (this study)
SCU893 mad2::kanM X [pMAD2-GFP] (this study)
SCU1338 (bub2Δ MAD2-GFP)
SCU151 mad2::kanMX [pMAD2-GFP] (this study)
SCU1700 (cdh1Δ CEN-GFP)
SCU1228 trp1::LacOx256:TRP1 his3::HIS3p-GFP13-LacI12NLS::HIS3 (this study)
SCU893 pds1::PDS1-HA3::URA3 (this study)
SCU2834 (bub2Δ cdh1Δ PDS1-HA3)
SCU1336 ura3 pds1::PDS1-HA3::URA3 (this study)
Plasmids used in this study
GAL1 URA3 CEN 
TRP1 CEN 
lacOx256 LEU2 integrative 
CUP1pro-GFP12-LacI12-NLS HIS3 integrative 
NLS-tetR-GFP LEU2 integrative 
tetO2x112 URA3 integrative 
NOP1-HA3GFP URA3 CEN (this study)
PDS1-HA3 URA3 integrative 
GAL-CDC14-His6 URA3 CEN 
GAL-SCC1-R180D/R268D-HA3 LEU2 integrative 
GAL-CDH1-GFP TRP1 CEN 
GAL1-BFA1 URA3 CEN 
MAD2-GFP URA3 CEN 
MET3-CDC20 TRP1 integrative (F. Uhlmann)
GAL1-PDS1 with mutated D-box URA3 CEN (this study)
GAL1-PDS1 with mutated D/KEN-box URA3 CEN (this study)
GAL1-MAD2-His6HAZZ 2 μ URA3 
GAL1-CDH1-TAP 2 μ TRP1 (this study)
GAL1-CDC14-His6HAZZ 2 μ TRP1 (this study)
MTW1-DsRed.T4 CEN LEU2 (this study)
Except for Mad2-GFP-expressing cells, cells expressing GFP-tagged proteins were fixed with 70% ethanol for 30 sec. After washing with distilled water, cells were stained with 4',6-diamidino-2-phenylindole (DAPI) at 1 μg/ml for 15 min. For detection of weak Mad2-GFP signals, cells were not fixed with ethanol. Washed cells were viewed using an Olympus IX71-23FL/S microscope (100× objective) and a cooled charge-couple device (CCD) camera (ORCA-ER-1, Hamamatsu Photonics) connected to a Scanalytics Image Processor LuminaVision (Mitani Corp., Tokyo, Japan). For Figures 4C and 5A and Additional file 1A, a Carl Zeiss Axio Imager M1 microscope with a cooled CCD camera (Carl Zeiss AxioCam MRm) was used.
Western blotting analysis
Western blotting was performed as described previously  using an anti-hemagglutinin (HA) antibody (16B12, BAbCo), anti-cyclin dependent kinase (CDK) antibody (Santa Cruz), and anti-glucose-6-phosphate dehydrogenase (G6PDH) antibody (Sigma). Femtogrow chemiluminescent substrate (Michigan Diagnostics) for horseradish peroxidase (HRP) and Can Get Signals (Toyobo, Japan) as an immunoreaction enhancer solution were used. Chemiluminescent signals were detected using an LAS3000 mini (Fuji).
Present address of M. Ueno: Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, 1-3-1 Kagamiyama, Higashi-Hiroshima 739-8530, Japan
We thank Andrew Murray, Kim Nasmyth, Frank Uhlmann, Orna Cohen-Fix, Matthias Peter, Leland Johnston, David Morgan, Uttam Surana, Kiwon Song, Booth Quimby and Benjamin Glick for generous gifts of materials, and Hisao Moriya and Kazunari Kaizu for discussion. This research was performed in part using an instrument at the Center for Instrumental Analysis of Shizuoka University. We especially thank laboratory members of TU for helpful discussion and support.
- Castro A, Bernis C, Vigneron S, Labbe JC, Lorca T: The anaphase-promoting complex: a key factor in the regulation of cell cycle. Oncogene 2005, 24: 314–325. 10.1038/sj.onc.1207973View ArticlePubMedGoogle Scholar
- Manchado E, Eguren M, Malumbres M: The anaphase-promoting complex/cyclosome (APC/C): cell-cycle-dependent and -independent functions. Biochem Soc Trans 2010, 38: 65–71. 10.1042/BST0380065View ArticlePubMedGoogle Scholar
- Peters JM: The anaphase promoting complex/cyclosome: a machine designed to destroy. Nat Rev Mol Cell Biol 2006, 7: 644–656. 10.1038/nrm1988View ArticlePubMedGoogle Scholar
- Pines J: Cubism and the cell cycle: the many faces of the APC/C. Nat Rev Mol Cell Biol 2011, 12: 427–438. 10.1038/nrm3132View ArticlePubMedGoogle Scholar
- Sullivan M, Morgan DO: Finishing mitosis, one step at a time. Nat Rev Mol Cell Biol 2007, 8: 894–903. 10.1038/nrm2276View ArticlePubMedGoogle Scholar
- Schwab M, Lutum AS, Seufert W: Yeast Hct1 is a regulator of Clb2 cyclin proteolysis. Cell 1997, 90: 683–693. 10.1016/S0092-8674(00)80529-2View ArticlePubMedGoogle Scholar
- Visintin R, Prinz S, Amon A: CDC20 and CDH1: a family of substrate-specific activators of APC-dependent proteolysis. Science 1997, 278: 460–463. 10.1126/science.278.5337.460View ArticlePubMedGoogle Scholar
- Simanis V: Events at the end of mitosis in the budding and fission yeasts. J Cell Sci 2003, 116: 4263–4275. 10.1242/jcs.00807View ArticlePubMedGoogle Scholar
- Stegmeier F, Amon A: Closing mitosis: the functions of the Cdc14 phosphatase and its regulation. Annu Rev Genet 2004, 38: 203–232. 10.1146/annurev.genet.38.072902.093051View ArticlePubMedGoogle Scholar
- Tan AL, Rida PC, Surana U: Essential tension and constructive destruction: the spindle checkpoint and its regulatory links with mitotic exit. Biochem J 2005, 386: 1–13. 10.1042/BJ20041415View ArticlePubMed CentralPubMedGoogle Scholar
- Bharadwaj R, Yu H: The spindle checkpoint, aneuploidy, and cancer. Oncogene 2004, 23: 2016–2027. 10.1038/sj.onc.1207374View ArticlePubMedGoogle Scholar
- Musacchio A, Salmon ED: The spindle-assembly checkpoint in space and time. Nat Rev Mol Cell Biol 2007, 8: 379–393. 10.1038/nrm2163View ArticlePubMedGoogle Scholar
- Kulukian A, Han JS, Cleveland DW: Unattached kinetochores catalyze production of an anaphase inhibitor that requires a Mad2 template to prime Cdc20 for BubR1 binding. Dev Cell 2009, 16: 105–117. 10.1016/j.devcel.2008.11.005View ArticlePubMed CentralPubMedGoogle Scholar
- Pinsky BA, Nelson CR, Biggins S: Protein phosphatase 1 regulates exit from the spindle checkpoint in budding yeast. Curr Biol 2009, 19: 1182–1187. 10.1016/j.cub.2009.06.043View ArticlePubMed CentralPubMedGoogle Scholar
- Goto GH, Mishra A, Abdulle R, Slaughter CA, Kitagawa K: Bub1-mediated adaptation of the spindle checkpoint. PLoS Genet 2011, 7: e1001282. 10.1371/journal.pgen.1001282View ArticlePubMed CentralPubMedGoogle Scholar
- Rieder CL, Maiato H: Stuck in division or passing through: what happens when cells cannot satisfy the spindle assembly checkpoint. Dev Cell 2004, 7: 637–651. 10.1016/j.devcel.2004.09.002View ArticlePubMedGoogle Scholar
- Minn AJ, Boise LH, Thompson CB: Expression of Bcl-xL and loss of p53 can cooperate to overcome a cell cycle checkpoint induced by mitotic spindle damage. Genes Dev 1996, 10: 2621–2631. 10.1101/gad.10.20.2621View ArticlePubMedGoogle Scholar
- Brito DA, Rieder CL: Mitotic checkpoint slippage in humans occurs via cyclin B destruction in the presence of an active checkpoint. Curr Biol 2006, 16: 1194–1200. 10.1016/j.cub.2006.04.043View ArticlePubMed CentralPubMedGoogle Scholar
- Gascoigne KE, Taylor SS: Cancer cells display profound intra- and interline variation following prolonged exposure to antimitotic drugs. Cancer Cell 2008, 14: 111–122. 10.1016/j.ccr.2008.07.002View ArticlePubMedGoogle Scholar
- Lee J, Kim JA, Margolis RL, Fotedar R: Substrate degradation by the anaphase promoting complex occurs during mitotic slippage. Cell Cycle 2010, 9: 1792–1801. 10.4161/cc.9.9.11519View ArticlePubMed CentralPubMedGoogle Scholar
- Fry AM, Yamano H: APC/C-mediated degradation in early mitosis: how to avoid spindle assembly checkpoint inhibition. Cell Cycle 2006, 5: 1487–1491. 10.4161/cc.5.14.3003View ArticlePubMedGoogle Scholar
- Hayes MJ, Kimata Y, Wattam SL, Lindon C, Mao G, Yamano H, Fry AM: Early mitotic degradation of Nek2A depends on Cdc20-independent interaction with the APC/C. Nat Cell Biol 2006, 8: 607–614. 10.1038/ncb1410View ArticlePubMedGoogle Scholar
- Rossio V, Galati E, Ferrari M, Pellicioli A, Sutani T, Shirahige K, Lucchini G, Piatti S: The RSC chromatin-remodeling complex influences mitotic exit and adaptation to the spindle assembly checkpoint by controlling the Cdc14 phosphatase. J Cell Biol 2010, 191: 981–997. 10.1083/jcb.201007025View ArticlePubMed CentralPubMedGoogle Scholar
- Hoyt MA, Totis L, Roberts BT: S. cerevisiae genes required for cell cycle arrest in response to loss of microtubule function. Cell 1991, 66: 507–517. 10.1016/0092-8674(81)90014-3View ArticlePubMedGoogle Scholar
- Bardin AJ, Amon A: Men and sin: what's the difference? Nat Rev Mol Cell Biol 2001, 2: 815–826.View ArticlePubMedGoogle Scholar
- Straight AF, Belmont AS, Robinett CC, Murray AW: GFP tagging of budding yeast chromosomes reveals that protein-protein interactions can mediate sister chromatid cohesion. Curr Biol 1996, 6: 1599–1608. 10.1016/S0960-9822(02)70783-5View ArticlePubMedGoogle Scholar
- Fraschini R, Formenti E, Lucchini G, Piatti S: Budding yeast Bub2 is localized at spindle pole bodies and activates the mitotic checkpoint via a different pathway from Mad2. J Cell Biol 1999, 145: 979–991. 10.1083/jcb.145.5.979View ArticlePubMed CentralPubMedGoogle Scholar
- Krishnan R, Pangilinan F, Lee C, Spencer F: Saccharomyces cerevisiae BUB2 prevents mitotic exit in response to both spindle and kinetochore damage. Genetics 2000, 156: 489–500.PubMed CentralPubMedGoogle Scholar
- Shirayama M, Zachariae W, Ciosk R, Nasmyth K: The Polo-like kinase Cdc5p and the WD-repeat protein Cdc20p/fizzy are regulators and substrates of the anaphase promoting complex in Saccharomyces cerevisiae . EMBO J 1998, 17: 1336–1349. 10.1093/emboj/17.5.1336View ArticlePubMed CentralPubMedGoogle Scholar
- Tavormina PA, Burke DJ: Cell cycle arrest in cdc20 mutants of Saccharomyces cerevisiae is independent of Ndc10p and kinetochore function but requires a subset of spindle checkpoint genes. Genetics 1998, 148: 1701–1713.PubMed CentralPubMedGoogle Scholar
- Cohen-Fix O, Peters JM, Kirschner MW, Koshland D: Anaphase initiation in Saccharomyces cerevisiae is controlled by the APC-dependent degradation of the anaphase inhibitor Pds1p. Genes Dev 1996, 10: 3081–3093. 10.1101/gad.10.24.3081View ArticlePubMedGoogle Scholar
- Holt LJ, Krutchinsky AN, Morgan DO: Positive feedback sharpens the anaphase switch. Nature 2008, 454: 353–357. 10.1038/nature07050View ArticlePubMed CentralPubMedGoogle Scholar
- Schwickart M, Havlis J, Habermann B, Bogdanova A, Camasses A, Oelschlaegel T, Shevchenko A, Zachariae W: Swm1/Apc13 is an evolutionarily conserved subunit of the anaphase-promoting complex stabilizing the association of Cdc16 and Cdc27. Mol Cell Biol 2004, 24: 3562–3576. 10.1128/MCB.24.8.3562-3576.2004View ArticlePubMed CentralPubMedGoogle Scholar
- Carroll CW, Enquist-Newman M, Morgan DO: The APC subunit Doc1 promotes recognition of the substrate destruction box. Curr Biol 2005, 15: 11–18. 10.1016/j.cub.2004.12.066View ArticlePubMedGoogle Scholar
- Nasmyth K: Segregating sister genomes: the molecular biology of chromosome separation. Science 2002, 297: 559–565. 10.1126/science.1074757View ArticlePubMedGoogle Scholar
- Meluh PB, Strunnikov AV: Beyond the ABCs of CKC and SCC. Do centromeres orchestrate sister chromatid cohesion or vice versa? Eur J Biochem 2002, 269: 2300–2314. 10.1046/j.1432-1033.2002.02886.xView ArticlePubMedGoogle Scholar
- Uhlmann F: Chromosome cohesion and separation: from men and molecules. Curr Biol 2003, 13: R104–114. 10.1016/S0960-9822(03)00039-3View ArticlePubMedGoogle Scholar
- Tang X, Wang Y: Pds1/Esp1-dependent and -independent sister chromatid separation in mutants defective for protein phosphatase 2A. Proc Natl Acad Sci USA 2006, 103: 16290–16295. 10.1073/pnas.0607856103View ArticlePubMed CentralPubMedGoogle Scholar
- Renshaw MJ, Ward JJ, Kanemaki M, Natsume K, Nedelec FJ, Tanaka TU: Condensins promote chromosome recoiling during early anaphase to complete sister chromatid separation. Dev Cell 2010, 19: 232–244. 10.1016/j.devcel.2010.07.013View ArticlePubMed CentralPubMedGoogle Scholar
- Uhlmann F, Lottspeich F, Nasmyth K: Sister-chromatid separation at anaphase onset is promoted by cleavage of the cohesin subunit Scc1. Nature 1999, 400: 37–42. 10.1038/21831View ArticlePubMedGoogle Scholar
- D'Amours D, Amon A: At the interface between signaling and executing anaphase--Cdc14 and the FEAR network. Genes Dev 2004, 18: 2581–2595. 10.1101/gad.1247304View ArticlePubMedGoogle Scholar
- Strunnikov AV: A case of selfish nucleolar segregation. Cell Cycle 2005, 4: 113–117. 10.4161/cc.4.1.1488View ArticlePubMedGoogle Scholar
- Torres-Rosell J, Machin F, Aragon L: Cdc14 and the temporal coordination between mitotic exit and chromosome segregation. Cell Cycle 2005, 4: 109–112. 10.4161/cc.4.1.1356View ArticlePubMedGoogle Scholar
- de Almeida A, Raccurt I, Peyrol S, Charbonneau M: The Saccharomyces cerevisiae Cdc14 phosphatase is implicated in the structural organization of the nucleolus. Biol Cell 1999, 91: 649–663.View ArticlePubMedGoogle Scholar
- Shirayama M, Toth A, Galova M, Nasmyth K: APC(Cdc20) promotes exit from mitosis by destroying the anaphase inhibitor Pds1 and cyclin Clb5. Nature 1999, 402: 203–207. 10.1038/46080View ArticlePubMedGoogle Scholar
- Pines J: Mitosis: a matter of getting rid of the right protein at the right time. Trends Cell Biol 2006, 16: 55–63. 10.1016/j.tcb.2005.11.006View ArticlePubMedGoogle Scholar
- van Leuken R, Clijsters L, Wolthuis R: To cell cycle, swing the APC/C. Biochim Biophys Acta 2008, 1786: 49–59.PubMedGoogle Scholar
- Wasch R, Robbins JA, Cross FR: The emerging role of APC/CCdh1 in controlling differentiation, genomic stability and tumor suppression. Oncogene 2010, 29: 1–10. 10.1038/onc.2009.325View ArticlePubMed CentralPubMedGoogle Scholar
- Rudner AD, Hardwick KG, Murray AW: Cdc28 activates exit from mitosis in budding yeast. J Cell Biol 2000, 149: 1361–1376. 10.1083/jcb.149.7.1361View ArticlePubMed CentralPubMedGoogle Scholar
- Stevenson LF, Kennedy BK, Harlow E: A large-scale overexpression screen in Saccharomyces cerevisiae identifies previously uncharacterized cell cycle genes. Proc Natl Acad Sci USA 2001, 98: 3946–3951. 10.1073/pnas.051013498View ArticlePubMed CentralPubMedGoogle Scholar
- Crasta K, Huang P, Morgan G, Winey M, Surana U: Cdk1 regulates centrosome separation by restraining proteolysis of microtubule-associated proteins. EMBO J 2006, 25: 2551–2563. 10.1038/sj.emboj.7601136View ArticlePubMed CentralPubMedGoogle Scholar
- Crasta K, Lim HH, Giddings TH Jr, Winey M, Surana U: Inactivation of Cdh1 by synergistic action of Cdk1 and polo kinase is necessary for proper assembly of the mitotic spindle. Nat Cell Biol 2008, 10: 665–675. 10.1038/ncb1729View ArticlePubMed CentralPubMedGoogle Scholar
- Krapp A, Gulli MP, Simanis V: SIN and the art of splitting the fission yeast cell. Curr Biol 2004, 14: R722–730. 10.1016/j.cub.2004.08.049View ArticlePubMedGoogle Scholar
- Krapp A, Simanis V: An overview of the fission yeast septation initiation network (SIN). Biochem Soc Trans 2008, 36: 411–415. 10.1042/BST0360411View ArticlePubMedGoogle Scholar
- Cerutti L, Simanis V: Asymmetry of the spindle pole bodies and spg1p GAP segregation during mitosis in fission yeast. J Cell Sci 1999, 112: 2313–2321.PubMedGoogle Scholar
- Mulvihill DP, Hyams JS: Cytokinetic actomyosin ring formation and septation in fission yeast are dependent on the full recruitment of the polo-like kinase Plo1 to the spindle pole body and a functional spindle assembly checkpoint. J Cell Sci 2002, 115: 3575–3586. 10.1242/jcs.00031View ArticlePubMedGoogle Scholar
- Liu J, Tang X, Wang H, Oliferenko S, Balasubramanian MK: The localization of the integral membrane protein Cps1p to the cell division site is dependent on the actomyosin ring and the septation-inducing network in Schizosaccharomyces pombe . Mol Biol Cell 2002, 13: 989–1000. 10.1091/mbc.01-12-0581View ArticlePubMed CentralPubMedGoogle Scholar
- Pesin JA, Orr-Weaver TL: Regulation of APC/C activators in mitosis and meiosis. Annu Rev Cell Dev Biol 2008, 24: 475–499. 10.1146/annurev.cellbio.041408.115949View ArticlePubMed CentralPubMedGoogle Scholar
- Zur A, Brandeis M: Securin degradation is mediated by fzy and fzr, and is required for complete chromatid separation but not for cytokinesis. EMBO J 2001, 20: 792–801. 10.1093/emboj/20.4.792View ArticlePubMed CentralPubMedGoogle Scholar
- Hagting A, Den Elzen N, Vodermaier HC, Waizenegger IC, Peters JM, Pines J: Human securin proteolysis is controlled by the spindle checkpoint and reveals when the APC/C switches from activation by Cdc20 to Cdh1. J Cell Biol 2002, 157: 1125–1137. 10.1083/jcb.200111001View ArticlePubMed CentralPubMedGoogle Scholar
- Jeganathan KB, Malureanu L, van Deursen JM: The Rae1-Nup98 complex prevents aneuploidy by inhibiting securin degradation. Nature 2005, 438: 1036–1039. 10.1038/nature04221View ArticlePubMedGoogle Scholar
- Honma Y, Kitamura A, Shioda R, Maruyama H, Ozaki K, Oda Y, Mini T, Jeno P, Maki Y, Yonezawa K, Hurt E, Ueno M, Uritani M, Hall MN, Ushimaru T: TOR regulates late steps of ribosome maturation in the nucleoplasm via Nog1 in response to nutrients. EMBO J 2006, 25: 3832–3842. 10.1038/sj.emboj.7601262View ArticlePubMed CentralPubMedGoogle Scholar
- Ross KE, Cohen-Fix O: The role of Cdh1p in maintaining genomic stability in budding yeast. Genetics 2003, 165: 489–503.PubMed CentralPubMedGoogle Scholar
- Mumberg D, Muller R, Funk M: Regulatable promoters of Saccharomyces cerevisiae : comparison of transcriptional activity and their use for heterologous expression. Nucleic Acids Res 1994, 22: 5767–5768. 10.1093/nar/22.25.5767View ArticlePubMed CentralPubMedGoogle Scholar
- Sikorski RS, Hieter P: A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae . Genetics 1989, 122: 19–27.PubMed CentralPubMedGoogle Scholar
- Michaelis C, Ciosk R, Nasmyth K: Cohesins: chromosomal proteins that prevent premature separation of sister chromatids. Cell 1997, 91: 35–45. 10.1016/S0092-8674(01)80007-6View ArticlePubMedGoogle Scholar
- Jensen S, Geymonat M, Johnson AL, Segal M, Johnston LH: Spatial regulation of the guanine nucleotide exchange factor Lte1 in Saccharomyces cerevisiae . J Cell Sci 2002, 115: 4977–4991. 10.1242/jcs.00189View ArticlePubMedGoogle Scholar
- Jaquenoud M, van Drogen F, Peter M: Cell cycle-dependent nuclear export of Cdh1p may contribute to the inactivation of APC/C(Cdh1). EMBO J 2002, 21: 6515–6526. 10.1093/emboj/cdf634View ArticlePubMed CentralPubMedGoogle Scholar
- Lee J, Hwang HS, Kim J, Song K: Ibd1p, a possible spindle pole body associated protein, regulates nuclear division and bud separation in Saccharomyces cerevisiae . Biochim Biophys Acta 1999, 1449: 239–253. 10.1016/S0167-4889(99)00015-4View ArticlePubMedGoogle Scholar
- Quimby BB, Arnaoutov A, Dasso M: Ran GTPase regulates Mad2 localization to the nuclear pore complex. Eukaryot Cell 2005, 4: 274–280. 10.1128/EC.4.2.274-280.2005View ArticlePubMed CentralPubMedGoogle Scholar
- Gelperin DM, White MA, Wilkinson ML, Kon Y, Kung LA, Wise KJ, Lopez-Hoyo N, Jiang L, Piccirillo SYuH, Gerstein M, Dumont ME, Phizicky EM, Snyder M, Grayhack EJ: Biochemical and genetic analysis of the yeast proteome with a movable ORF collection. Genes Dev 2005, 19: 2816–2826. 10.1101/gad.1362105View ArticlePubMed CentralPubMedGoogle Scholar
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