Farnesol-induced growth inhibition in Saccharomyces cerevisiae by a cell cycle mechanism

Heterologous expression of bacterial virulence factors in Saccharomyces cerevisiae is a feasible approach to study their molecular function. The authors have previously reported that the Salmonella typhimurium SigD protein, a phosphatidylinositol phosphatase involved in invasion of the host cell, inhibits yeast growth, presumably by depleting an essential pool of phosphatidylinositol 4,5-bisphosphate, and also that a catalytically inactive version, SigDR468A, was able to arrest growth by a different mechanism that involved disruption of the actin cytoskeleton. This paper describes marked differences between the phenotypes elicited by expression of SigD and SigDR468A in yeast. First, expression of SigDR468A caused accumulation of large unbudded cells and loss of septin organization, while SigD expression caused none of these effects. Second, growth inhibition by SigDR468A was mediated by a cell cycle arrest in G2 dependent on the Swe1 morphogenetic checkpoint, but SigD-induced growth inhibition was cell cycle independent. And third, SigD caused strong activation of the yeast MAP kinase Slt2, whereas SigDR468A rather inactivated another MAP kinase, Kss1. In a screen for suppressors of SigDR468A-induced growth arrest by overexpression of a yeast cDNA library, the Cdc42 GTPase was isolated. Furthermore, SigDR468A was co-purified with Cdc42 from yeast lysates. It is concluded that the Salmonella SigD protein deprived of its phosphatase activity is able to disrupt yeast morphogenesis by interfering with Cdc42 function, opening the possibility that the SigD N-terminal region might directly modulate small GTPases from the host during infection.

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Chs1p and Chs3p, two proteins involved in chitin synthesis, populate a compartment of the Saccharomyces cerevisiae endocytic pathway.

Molecular Biology of the Cell ◽

10.1091/mbc.7.12.1909 ◽

1996 ◽

Vol 7 (12) ◽

pp. 1909-1919 ◽

Cited By ~ 104

Author(s):

M Ziman ◽

J S Chuang ◽

R W Schekman

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Endocytic Pathway ◽

Chitin Synthase ◽

Cell Fractionation ◽

Cell Wall Polysaccharide ◽

Chitin Synthesis ◽

A Cell ◽

Steady State Levels ◽

Membrane Bound

In Saccharomyces cerevisiae, the synthesis of chitin, a cell-wall polysaccharide, is temporally and spatially regulated with respect to the cell cycle and morphogenesis. Using immunological reagents, we found that steady-state levels of Chs1p and Chs3p, two chitin synthase enzymes, did not fluctuate during the cell cycle, indicating that they are not simply regulated by synthesis and degradation. Previous cell fractionation studies demonstrated that chitin synthase I activity (CSI) exists in a plasma membrane form and in intracellular membrane-bound particles called chitosomes. Chitosomes were proposed to act as a reservoir for regulated transport of chitin synthase enzymes to the division septum. We found that Chs1p and Chs3p resided partly in chitosomes and that this distribution was not cell cycle regulated. Pulse-chase cell fractionation experiments showed that chitosome production was blocked in an endocytosis mutant (end4-1), indicating that endocytosis is required for the formation or maintenance of chitosomes. Additionally, Ste2p, internalized by ligand-induced endocytosis, cofractionated with chitosomes, suggesting that these membrane proteins populate the same endosomal compartment. However, in contrast to Ste2p, Chs1p and Chs3p were not rapidly degraded, thus raising the possibility that the temporal and spatial regulation of chitin synthesis is mediated by the mobilization of an endosomal pool of chitin synthase enzymes.

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Cell cycle–dependent phosphorylation of Sec4p controls membrane deposition during cytokinesis

The Journal of Cell Biology ◽

10.1083/jcb.201602038 ◽

2016 ◽

Vol 214 (6) ◽

pp. 691-703 ◽

Cited By ~ 9

Author(s):

Dante Lepore ◽

Olya Spassibojko ◽

Gabrielle Pinto ◽

Ruth N. Collins

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Membrane Trafficking ◽

Intracellular Trafficking ◽

Rab Gtpase ◽

Rab Gtpases ◽

Positive Regulator ◽

Physiological Relevance ◽

A Cell ◽

Cycle Dependent

Intracellular trafficking is an essential and conserved eukaryotic process. Rab GTPases are a family of proteins that regulate and provide specificity for discrete membrane trafficking steps by harnessing a nucleotide-bound cycle. Global proteomic screens have revealed many Rab GTPases as phosphoproteins, but the effects of this modification are not well understood. Using the Saccharomyces cerevisiae Rab GTPase Sec4p as a model, we have found that phosphorylation negatively regulates Sec4p function by disrupting the interaction with the exocyst complex via Sec15p. We demonstrate that phosphorylation of Sec4p is a cell cycle–dependent process associated with cytokinesis. Through a genomic kinase screen, we have also identified the polo-like kinase Cdc5p as a positive regulator of Sec4p phosphorylation. Sec4p spatially and temporally localizes with Cdc5p exclusively when Sec4p phosphorylation levels peak during the cell cycle, indicating Sec4p is a direct Cdc5p substrate. Our data suggest the physiological relevance of Sec4p phosphorylation is to facilitate the coordination of membrane-trafficking events during cytokinesis.

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Saccharomyces cerevisiae cdc15 mutants arrested at a late stage in anaphase are rescued by Xenopus cDNAs encoding N-ras or a protein with beta-transducin repeats.

Molecular and Cellular Biology ◽

10.1128/mcb.13.8.4953 ◽

1993 ◽

Vol 13 (8) ◽

pp. 4953-4966 ◽

Cited By ~ 44

Author(s):

W Spevak ◽

B D Keiper ◽

C Stratowa ◽

M J Castañón

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Yeast Cells ◽

Cell Cycle Gene ◽

Intracellular Camp ◽

Temperature Sensitive ◽

Ras Genes ◽

Phosphorylation Event ◽

Dependent Protein ◽

A Cell

We have constructed a Xenopus oocyte cDNA library in a Saccharomyces cerevisiae expression vector and used this library to isolate genes that can function in yeast cells to suppress the temperature sensitive [corrected] defect of the cdc15 mutation. Two maternally expressed Xenopus cDNAs which fulfill these conditions have been isolated. One of these clones encodes Xenopus N-ras. In contrast to the yeast RAS genes, Xenopus N-ras rescues the cdc15 mutation. Moreover, overexpression of Xenopus N-ras in S. cerevisiae does not activate the RAS-cyclic AMP (cAMP) pathway; rather, it results in decreased levels of intracellular cAMP in both mutant cdc15 and wild-type cells. Furthermore, we show that lowering cAMP levels is sufficient to allow cells with a nonfunctional Cdc15 protein to complete the mitotic cycle. These results suggest that a key step of the cell cycle is dependent upon a phosphorylation event catalyzed by cAMP-dependent protein kinase. The second clone, beta TrCP (beta-transducin repeat-containing protein), encodes a protein of 518 amino acids that shows significant homology to the beta subunits of G proteins in its C-terminal half. In this region, beta Trcp is composed of seven beta-transducin repeats. beta TrCP is not a functional homolog of S. cerevisiae CDC20, a cell cycle gene that also contains beta-transducin repeats and suppresses the cdc15 mutation.

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The Saccharomyces cerevisiae Telomerase Subunit Est3 Binds Telomeres in a Cell Cycle– and Est1–Dependent Manner and Interacts Directly with Est1 In Vitro

PLoS Genetics ◽

10.1371/journal.pgen.1002060 ◽

2011 ◽

Vol 7 (5) ◽

pp. e1002060 ◽

Cited By ~ 28

Author(s):

Creighton T. Tuzon ◽

Yun Wu ◽

Angela Chan ◽

Virginia A. Zakian

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Dependent Manner ◽

A Cell

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Dpb11, which interacts with DNA polymerase II(epsilon) in Saccharomyces cerevisiae, has a dual role in S-phase progression and at a cell cycle checkpoint.

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.92.25.11791 ◽

1995 ◽

Vol 92 (25) ◽

pp. 11791-11795 ◽

Cited By ~ 165

Author(s):

H. Araki ◽

S. H. Leem ◽

A. Phongdara ◽

A. Sugino

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Dna Polymerase ◽

S Phase ◽

Dual Role ◽

Cell Cycle Checkpoint ◽

Cycle Checkpoint ◽

A Cell ◽

Phase Progression ◽

Dna Polymerase Ii

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The Saccharomyces cerevisiae RNase Mitochondrial RNA Processing Is Critical for Cell Cycle Progression at the End of Mitosis

Genetics ◽

10.1093/genetics/161.3.1029 ◽

2002 ◽

Vol 161 (3) ◽

pp. 1029-1042 ◽

Cited By ~ 1

Author(s):

Ti Cai ◽

Jason Aulds ◽

Tina Gill ◽

Michael Cerio ◽

Mark E Schmitt

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Cell Cycle Progression ◽

Mitochondrial Rna ◽

Mrna Levels ◽

Cycle Progression ◽

Rnase Mrp ◽

Processing Step ◽

A Cell

Abstract We have identified a cell cycle delay in Saccharomyces cerevisiae RNase MRP mutants. Mutants delay with large budded cells, dumbbell-shaped nuclei, and extended spindles characteristic of “exit from mitosis” mutants. In accord with this, a RNase MRP mutation can be suppressed by overexpressing the polo-like kinase CDC5 or by deleting the B-type cyclin CLB1, without restoring the MRP-dependent rRNA-processing step. In addition, we identified a series of genetic interactions between RNase MRP mutations and mutations in CDC5, CDC14, CDC15, CLB2, and CLB5. As in most “exit from mitosis” mutants, levels of the Clb2 cyclin were increased. The buildup of Clb2 protein is not the result of a defect in the release of the Cdc14 phosphatase from the nucleolus, but rather the result of an increase in CLB2 mRNA levels. These results indicate a clear role of RNase MRP in cell cycle progression at the end of mitosis. Conservation of this function in humans may explain many of the pleiotropic phenotypes of cartilage hair hypoplasia.

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LncRNAs of Saccharomyces cerevisiae dodge the cell cycle arrest imposed by the ethanol stress

10.1101/2021.06.28.450142 ◽

2021 ◽

Author(s):

Lucas Cardoso Lázari ◽

Ivan Rodrigo Wolf ◽

Amanda Piveta Schnepper ◽

Guilherme Targino Valente

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Cell Cycle Arrest ◽

Ethanol Stress ◽

Network Dynamic ◽

Cell Cycle Proteins ◽

Tolerant Strain ◽

Cycle Arrest ◽

A Cell ◽

The One

Ethanol impairs many subsystems of Saccharomyces cerevisiae, including the cell cycle. Cyclins and damage checkpoints drive the cell cycle. Two ethanol-responsive lncRNAs in yeast interact with cell cycle proteins, and here we investigated the role of these RNAs on the ethanol-stressed cell cycle. Our network dynamic modeling showed that the higher and lower ethanol tolerant strains undergo a cell cycle arrest during the ethanol stress. However, lower tolerant phenotype arrest in a later phase leading to its faster population rebound after the stress relief. Two lncRNAs can skip the arrests mentioned. The in silico overexpression of lnc9136 of SEY6210 (a lower tolerant strain), and CRISPR-Cas9 partial deletions of this lncRNA, evidenced that the one induces a regular cell cycle even under ethanol stress; this lncRNA binds to Gin4 and Hsl1, driving the Swe1p, Clb1/2, and cell cycle. Moreover, the lnc10883 of BY4742 (a higher tolerant strain) interacts to the Mec1p and represses Bub1p, circumventing the DNA and spindle damage checkpoints keeping a normal cell cycle even under DNA damage. Overall, we present the first evidence of the direct roles of lncRNAs on cell cycle proteins, the dynamics of this system in different ethanol tolerant phenotypes, and a new cell cycle model.

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Interaction between the plant ApDef1 defensin and Saccharomyces cerevisiae results in yeast death through a cell cycle- and caspase-dependent process occurring via uncontrolled oxidative stress

Biochimica et Biophysica Acta (BBA) - General Subjects ◽

10.1016/j.bbagen.2016.09.005 ◽

2017 ◽

Vol 1861 (1) ◽

pp. 3429-3443 ◽

Cited By ~ 13

Author(s):

Júlia Ribeiro Soares ◽

Edésio José Tenório de Melo ◽

Maura da Cunha ◽

Kátia Valevski Sales Fernandes ◽

Gabriel Bonan Taveira ◽

...

Keyword(s):

Oxidative Stress ◽

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Dependent Process ◽

A Cell

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SLA2 mutations cause SWE1-mediated cell cycle phenotypes in Candida albicans and Saccharomyces cerevisiae

Microbiology ◽

10.1099/mic.0.033233-0 ◽

2009 ◽

Vol 155 (12) ◽

pp. 3847-3859 ◽

Cited By ~ 12

Author(s):

Cheryl A. Gale ◽

Michelle D. Leonard ◽

Kenneth R. Finley ◽

Leah Christensen ◽

Mark McClellan ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Candida Albicans ◽

Actin Organization ◽

Growth Defects ◽

A Cell ◽

Morphogenesis Checkpoint ◽

Actin Patch ◽

Actin Cables ◽

Cell Cycle Dynamics

The early endocytic patch protein Sla2 is important for morphogenesis and growth rates in Saccharomyces cerevisiae and Candida albicans, but the mechanism that connects these processes is not clear. Here we report that growth defects in cells lacking CaSLA2 or ScSLA2 are associated with a cell cycle delay that is influenced by Swe1, a morphogenesis checkpoint kinase. To establish how Swe1 monitors Sla2 function, we compared actin organization and cell cycle dynamics in strains lacking other components of early endocytic patches (Sla1 and Abp1) with those in strains lacking Sla2. Only sla2 strains had defects in actin cables, a known trigger of the morphogenesis checkpoint, yet all three strains exhibited Swe1-dependent phenotypes. Thus, Swe1 appears to monitor actin patch in addition to actin cable function. Furthermore, Swe1 contributed to virulence in a mouse model of disseminated candidiasis, implying a role for the morphogenesis checkpoint during the pathogenesis of C. albicans infections.

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