Aurora promotes cell division during recovery from TOR-mediated cell cycle arrest by driving spindle pole body recruitment of Polo

Centrosomes play critical roles in the cell division cycle and ciliogenesis. Sfi1 is a centrin-binding protein conserved from yeast to humans. Budding yeast Sfi1 is essential for the initiation of spindle pole body (SPB; yeast centrosome) duplication. However, the recruitment and partitioning of Sfi1 to centrosomal structures have never been fully investigated in any organism, and the presumed importance of the conserved tryptophans in the internal repeats of Sfi1 remains untested. Here we report that in fission yeast, instead of doubling abruptly at the initiation of SPB duplication and remaining at a constant level thereafter, Sfi1 is gradually recruited to SPBs throughout the cell cycle. Like an sfi1Δ mutant, a Trp-to-Arg mutant (sfi1-M46) forms monopolar spindles and exhibits mitosis and cytokinesis defects. Sfi1-M46 protein associates preferentially with one of the two daughter SPBs during mitosis, resulting in a failure of new SPB assembly in the SPB receiving insufficient Sfi1. Although all five conserved tryptophans tested are involved in Sfi1 partitioning, the importance of the individual repeats in Sfi1 differs. In summary, our results reveal a link between the conserved tryptophans and Sfi1 partitioning and suggest a revision of the model for SPB assembly.

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Functional Interaction Between the PKC1 Pathway and CDC31 Network of SPB Duplication Genes

Genetics ◽

10.1093/genetics/155.4.1543 ◽

2000 ◽

Vol 155 (4) ◽

pp. 1543-1559 ◽

Cited By ~ 3

Author(s):

Waheeda Khalfan ◽

Irena Ivanovska ◽

Mark D Rose

Keyword(s):

Cell Cycle ◽

Cell Wall ◽

Spindle Pole Body ◽

Spindle Pole ◽

Genetic Interactions ◽

Genetic Screens ◽

Functional Interaction ◽

Cycle Arrest ◽

Pole Body ◽

Synthetically Lethal

Abstract The earliest known step in yeast spindle pole body (SPB) duplication requires Cdc31p and Kar1p, two physically interacting SPB components, and Dsk2p and Rad23p, a pair of ubiquitin-like proteins. Components of the PKC1 pathway were found to interact with these SPB duplication genes in two independent genetic screens. Initially, SLG1 and PKC1 were obtained as high-copy suppressors of dsk2Δ rad23Δ and a mutation in MPK1 was synthetically lethal with kar1-Δ17. Subsequently, we demonstrated extensive genetic interactions between the PKC1 pathway and the SPB duplication mutants that affect Cdc31p function. The genetic interactions are unlikely to be related to the cell-wall integrity function of the PKC1 pathway because the SPB mutants did not exhibit cell-wall defects. Overexpression of multiple PKC1 pathway components suppressed the G2/M arrest of the SPB duplication mutants and mutations in MPK1 exacerbated the cell cycle arrest of kar1-Δ17, suggesting a role for the PKC1 pathway in SPB duplication. We also found that mutations in SPC110, which encodes a major SPB component, showed genetic interactions with both CDC31 and the PKC1 pathway. In support of the model that the PKC1 pathway regulates SPB duplication, one of the phosphorylated forms of Spc110p was absent in pkc1 and mpk1Δ mutants.

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Translational control of MPS1 links protein synthesis with the initiation of cell division and spindle pole body duplication in yeast

10.1101/2020.12.29.424704 ◽

2020 ◽

Author(s):

Heidi M. Blank ◽

Annabel Alonso ◽

Mark Winey ◽

Michael Polymenis

Keyword(s):

Cell Cycle ◽

Protein Synthesis ◽

Cell Division ◽

Translational Control ◽

Spindle Pole Body ◽

Spindle Pole ◽

Microtubule Organizing Center ◽

Structured Illumination Microscopy ◽

Couple Growth ◽

Pole Body

ABSTRACTProtein synthesis underpins cell growth and controls when cells commit to a new round of cell division at a point in late G1 of the cell cycle called Start. Passage through Start also coincides with the duplication of the microtubule-organizing centers, the yeast spindle pole bodies, which will form the two poles of the mitotic spindle that segregates the chromosomes in mitosis. The conserved Mps1p kinase governs the duplication of the spindle pole body in Saccharomyces cerevisiae. Here, we show that the MPS1 transcript has a short upstream open reading frame that represses the synthesis of Mps1p. Mutating the MPS1 uORF makes the cells smaller, accelerates the appearance of Mps1p in late G1, and promotes completion of Start. The accelerated Start of MPS1 uORF mutants depends on the G1 cyclin Cln3p. Monitoring the spindle pole body in the cell cycle using structured illumination microscopy revealed that mutating the MPS1 uORF enabled cells to duplicate their spindle pole body earlier, at a smaller cell size. For the first time, these results identify growth inputs in mechanisms that control duplication of the microtubule-organizing center and implicate these processes in general mechanisms that couple growth with division.

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Anticancer potential of nitric oxide (NO) in neuroblastoma treatment

RSC Advances ◽

10.1039/d1ra00275a ◽

2021 ◽

Vol 11 (16) ◽

pp. 9112-9120

Author(s):

Jenna L. Gordon ◽

Kristin J. Hinsen ◽

Melissa M. Reynolds ◽

Tyler A. Smith ◽

Haley O. Tucker ◽

...

Keyword(s):

Nitric Oxide ◽

Cell Cycle ◽

Cell Division ◽

Cell Viability ◽

Cell Cycle Arrest ◽

Neuroblastoma Cells ◽

Cycle Arrest

S-Nitrosoglutathione (GSNO) reduces cell viability, inhibits cell division, and induces cell cycle arrest and apoptosis in neuroblastoma cells.

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A novel small-molecule fatty acid synthase inhibitor with antitumor activity by cell cycle arrest and cell division inhibition

European Journal of Medicinal Chemistry ◽

10.1016/j.ejmech.2021.113407 ◽

2021 ◽

pp. 113407

Author(s):

Hongyan Qu ◽

Kai Shan ◽

Chunlei Tang ◽

Guozhen Cui ◽

Guoling Fu ◽

...

Keyword(s):

Cell Cycle ◽

Fatty Acid ◽

Cell Division ◽

Antitumor Activity ◽

Cell Cycle Arrest ◽

Small Molecule ◽

Fatty Acid Synthase ◽

Cycle Arrest ◽

Synthase Inhibitor ◽

Cell Division Inhibition

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Studies concerning the temporal and genetic control of cell polarity in Saccharomyces cerevisiae.

The Journal of Cell Biology ◽

10.1083/jcb.114.3.515 ◽

1991 ◽

Vol 114 (3) ◽

pp. 515-532 ◽

Cited By ~ 78

Author(s):

M Snyder ◽

S Gehrung ◽

B D Page

Keyword(s):

Cell Cycle ◽

Cell Polarity ◽

Spindle Pole Body ◽

Spindle Pole ◽

Yeast Cells ◽

Cell Cycle Distribution ◽

Restrictive Temperature ◽

Microtubule Bundle ◽

Pole Body ◽

Capture Site

The establishment of cell polarity was examined in the budding yeast, S. cerevisiae. The distribution of a polarized protein, the SPA2 protein, was followed throughout the yeast cell cycle using synchronized cells and cdc mutants. The SPA2 protein localizes to a patch at the presumptive bud site of G1 cells. Later it concentrates at the bud tip in budded cells. At cytokinesis, the SPA2 protein is at the neck between the mother and daughter cells. Analysis of unbudded haploid cells has suggested a series of events that occurs during G1. The SPA2 patch is established very early in G1, while the spindle pole body residues on the distal side of the nucleus. Later, microtubules emanating from the spindle pole body intersect the SPA2 crescent, and the nucleus probably rotates towards the SPA2 patch. By middle G1, most cells contain the SPB on the side of the nucleus proximal to the SPA2 patch, and a long extranuclear microtubule bundle intersects this patch. We suggest that a microtubule capture site exists in the SPA2 staining region that stabilizes the long microtubule bundle; this capture site may be responsible for rotation of the nucleus. Cells containing a polarized distribution of the SPA2 protein also possess a polarized distribution of actin spots in the same region, although the actin staining is much more diffuse. Moreover, cdc4 mutants, which form multiple buds at the restrictive temperature, exhibit simultaneous staining of the SPA2 protein and actin spots in a subset of the bud tips. spa2 mutants contain a polarized distribution of actin spots, and act1-1 and act1-2 mutants often contain a polarized distribution of the SPA2 protein suggesting that the SPA2 protein is not required for localization of the actin spots and the actin spots are not required for localization of the SPA2 protein. cdc24 mutants, which fail to form buds at the restrictive temperature, fail to exhibit polarized localization of the SPA2 protein and actin spots, indicating that the CDC24 protein is directly or indirectly responsible for controlling the polarity of these proteins. Based on the cell cycle distribution of the SPA2 protein, a "cytokinesis tag" model is proposed to explain the mechanism of the non-random positioning of bud sites in haploid yeast cells.

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Cdk1 promotes cytokinesis in fission yeast through activation of the septation initiation network

Molecular Biology of the Cell ◽

10.1091/mbc.e14-04-0936 ◽

2014 ◽

Vol 25 (15) ◽

pp. 2250-2259 ◽

Cited By ~ 6

Author(s):

Nicole Rachfall ◽

Alyssa E. Johnson ◽

Sapna Mehta ◽

Jun-Song Chen ◽

Kathleen L. Gould

Keyword(s):

Cell Cycle ◽

Spindle Pole Body ◽

Complete Removal ◽

Spindle Pole ◽

Cyclin Dependent Kinase ◽

Gtpase Activating Protein ◽

Pole Body ◽

Kinase Cascade ◽

Septation Initiation Network ◽

Cycle Dependent

In Schizosaccharomyces pombe, late mitotic events are coordinated with cytokinesis by the septation initiation network (SIN), an essential spindle pole body (SPB)–associated kinase cascade, which controls the formation, maintenance, and constriction of the cytokinetic ring. It is not fully understood how SIN initiation is temporally regulated, but it depends on the activation of the GTPase Spg1, which is inhibited during interphase by the essential bipartite GTPase-activating protein Byr4-Cdc16. Cells are particularly sensitive to the modulation of Byr4, which undergoes cell cycle–dependent phosphorylation presumed to regulate its function. Polo-like kinase, which promotes SIN activation, is partially responsible for Byr4 phosphorylation. Here we show that Byr4 is also controlled by cyclin-dependent kinase (Cdk1)–mediated phosphorylation. A Cdk1 nonphosphorylatable Byr4 phosphomutant displays severe cell division defects, including the formation of elongated, multinucleate cells, failure to maintain the cytokinetic ring, and compromised SPB association of the SIN kinase Cdc7. Our analyses show that Cdk1-mediated phosphoregulation of Byr4 facilitates complete removal of Byr4 from metaphase SPBs in concert with Plo1, revealing an unexpected role for Cdk1 in promoting cytokinesis through activation of the SIN pathway.

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The influence of the microtubule inhibitor, methyl benzimidazol-2-yl-carbamate (MBC) on nuclear division and the cell cycle in Saccharomyces cerevisiae

Journal of Cell Science ◽

10.1242/jcs.46.1.341 ◽

1980 ◽

Vol 46 (1) ◽

pp. 341-352

Author(s):

R.A. Quinlan ◽

C.I. Pogson ◽

K. Gull

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Spindle Pole Body ◽

Spindle Pole ◽

Ultrastructural Examination ◽

Microtubule Inhibitor ◽

Microtubule Polymerization ◽

Pole Body ◽

Outer Component ◽

Spindle Microtubules

Methyl benzimidazol-2-yl-carbamate (MBC), at a concentration of 100 microM, has a pronounced effect on the growth of Saccharomyces cerevisiae, resulting in the accumulation of cells as large doublets. We have determined a specific execution point for the effect of MBC on the yeast cell cycle, and have shown that this execution point is between the cycle events of spindle pole body duplication and spindle pole body separation. An ultrastructural examination of the MBC-treated cells revealed the absence of cytoplasmic and spindle microtubules. MBC treatment also produced an altered spindle pole body morphology, causing the disappearance of the outer component. Nuclear size was also markedly increased in the MBC-induced doublet cells, although the septa were completely absent from these doublet cells. It is proposed that MBC inhibits microtubule polymerization, rather than causing the depolymerization of stable microtubules.

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Spc98p Directs the Yeast γ-Tubulin Complex into the Nucleus and Is Subject to Cell Cycle-dependent Phosphorylation on the Nuclear Side of the Spindle Pole Body

Molecular Biology of the Cell ◽

10.1091/mbc.9.4.775 ◽

1998 ◽

Vol 9 (4) ◽

pp. 775-793 ◽

Cited By ~ 63

Author(s):

Gislene Pereira ◽

Michael Knop ◽

Elmar Schiebel

Keyword(s):

Cell Cycle ◽

Spindle Pole Body ◽

Mitotic Checkpoint ◽

Spindle Pole ◽

Nuclear Localization Sequence ◽

Checkpoint Control ◽

Yeast Saccharomyces Cerevisiae ◽

Pole Body ◽

Cycle Dependent ◽

Cytoplasmic Side

In the yeast Saccharomyces cerevisiae, microtubules are organized by the spindle pole body (SPB), which is embedded in the nuclear envelope. Microtubule organization requires the γ-tubulin complex containing the γ-tubulin Tub4p, Spc98p, and Spc97p. The Tub4p complex is associated with cytoplasmic and nuclear substructures of the SPB, which organize the cytoplasmic and nuclear microtubules. Here we present evidence that the Tub4p complex assembles in the cytoplasm and then either binds to the cytoplasmic side of the SPB or is imported into the nucleus followed by binding to the nuclear side of the SPB. Nuclear import of the Tub4p complex is mediated by the essential nuclear localization sequence of Spc98p. Our studies also indicate that Spc98p in the Tub4p complex is phosphorylated at the nuclear, but not at the cytoplasmic, side of the SPB. This phosphorylation is cell cycle dependent and occurs after SPB duplication and nucleation of microtubules by the new SPB and therefore may have a role in mitotic spindle function. In addition, activation of the mitotic checkpoint stimulates Spc98p phosphorylation. The kinase Mps1p, which functions in SPB duplication and mitotic checkpoint control, seems to be involved in Spc98p phosphorylation. Our results also suggest that the nuclear and cytoplasmic Tub4p complexes are regulated differently.

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