Induction of a Mitosis Delay and Cell Lysis by High-Level Secretion of Mouse α-Amylase from Saccharomyces cerevisiae

ABSTRACT Some foreign proteins are produced in yeast in a cell cycle-dependent manner, but the cause of the cell cycle dependency is unknown. In this study, we found that Saccharomyces cerevisiae cells secreting high levels of mouse α-amylase have elongated buds and are delayed in cell cycle completion in mitosis. The delayed cell mitosis suggests that critical events during exit from mitosis might be disturbed. We found that the activities of PP2A (protein phosphatase 2A) and MPF (maturation-promoting factor) were reduced in α-amylase-oversecreting cells and that these cells showed a reduced level of assembly checkpoint protein Cdc55, compared to the accumulation in wild-type cells. MPF inactivation is due to inhibitory phosphorylation on Cdc28, as a cdc28mutant which lacks an inhibitory phosphorylation site on Cdc28 prevents MPF inactivation and prevents the defective bud morphology induced by overproduction of α-amylase. Our data also suggest that high levels of α-amylase may downregulate PPH22,leading to cell lysis. In conclusion, overproduction of heterologous α-amylase in S. cerevisiaeresults in a negative regulation of PP2A, which causes mitotic delay and leads to cell lysis.

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Truncated APC regulates the transcriptional activity of β-catenin in a cell cycle dependent manner

Human Molecular Genetics ◽

10.1093/hmg/ddl464 ◽

2006 ◽

Vol 16 (2) ◽

pp. 199-209 ◽

Cited By ~ 38

Author(s):

Jean Schneikert ◽

Annette Grohmann ◽

Jürgen Behrens

Keyword(s):

Cell Cycle ◽

Transcriptional Activity ◽

Dependent Manner ◽

A Cell ◽

Cycle Dependent

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Ser68 phosphoregulation is essential for CENP-A deposition, centromere function and viability in mice

10.1101/2021.11.23.469796 ◽

2021 ◽

Author(s):

Yuting Liu ◽

Kehui Wang ◽

Li Huang ◽

Jicheng Zhao ◽

Xinpeng Chen ◽

...

Keyword(s):

Cell Cycle ◽

Cell Viability ◽

Histone H3 ◽

Dependent Manner ◽

Centromere Function ◽

Histone H3 Variant ◽

A Cell ◽

Cycle Dependent ◽

Spatiotemporal Control

Centromere identity is defined by nucleosomes containing CENP-A, a histone H3 variant. The deposition of CENP-A at centromeres is tightly regulated in a cell-cycle-dependent manner. We previously reported that the spatiotemporal control of centromeric CENP-A incorporation is mediated by the phosphorylation of CENP-A Ser68. However, a recent report argued that Ser68 phosphoregulation is dispensable for accurate CENP-A loading. Here, we report that the substitution of Ser68 of endogenous CENP-A with either Gln68 or Glu68 severely impairs CENP-A deposition and cell viability. We also find that mice harboring the corresponding mutations are lethal. Together, these results indicate that the dynamic phosphorylation of Ser68 ensures cell-cycle-dependent CENP-A deposition and cell viability.

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Gcn5-mediated acetylation at MBF-regulated promoters induces the G1/S transcriptional wave

Nucleic Acids Research ◽

10.1093/nar/gkz561 ◽

2019 ◽

Vol 47 (16) ◽

pp. 8439-8451 ◽

Cited By ~ 1

Author(s):

Alberto González-Medina ◽

Elena Hidalgo ◽

José Ayté

Keyword(s):

Cell Cycle ◽

Molecular Mechanisms ◽

Histone H3 ◽

Dependent Manner ◽

Saga Complex ◽

Lysine Residues ◽

Physical Barriers ◽

A Cell ◽

Cycle Dependent ◽

Regulated Promoters

Abstract In fission yeast, MBF-dependent transcription is inactivated at the end of S phase through a negative feedback loop that involves the co-repressors, Yox1 and Nrm1. Although this repression system is well known, the molecular mechanisms involved in MBF activation remain largely unknown. Compacted chromatin constitutes a barrier to activators accessing promoters. Here, we show that chromatin regulation plays a key role in activating MBF-dependent transcription. Gcn5, a part of the SAGA complex, binds to MBF-regulated promoters through the MBF co-activator Rep2 in a cell cycle-dependent manner and in a reverse correlation to the binding of the MBF co-repressors, Nrm1 or Yox1. We propose that the co-repressors function as physical barriers to SAGA recruitment onto MBF promoters. We also show that Gcn5 acetylates specific lysine residues on histone H3 in a cell cycle-regulated manner. Furthermore, either in a gcn5 mutant or in a strain in which histone H3 is kept in an unacetylated form, MBF-dependent transcription is downregulated. In summary, Gcn5 is required for the full activation and correct timing of MBF-regulated gene transcription.

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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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Correction: Optineurin Regulates the Interferon Response in a Cell Cycle-Dependent Manner

PLoS Pathogens ◽

10.1371/journal.ppat.1004971 ◽

2015 ◽

Vol 11 (6) ◽

pp. e1004971 ◽

Cited By ~ 5

Author(s):

Pierre Génin ◽

Frédérique Cuvelier ◽

Sandrine Lambin ◽

Josina Côrte-Real Filipe ◽

Elodie Autrusseau ◽

...

Keyword(s):

Cell Cycle ◽

Interferon Response ◽

Dependent Manner ◽

A Cell ◽

Cycle Dependent

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Coupling Phosphate Homeostasis to Cell Cycle-Specific Transcription: Mitotic Activation of Saccharomyces cerevisiae PHO5 by Mcm1 and Forkhead Proteins

Molecular and Cellular Biology ◽

10.1128/mcb.00222-09 ◽

2009 ◽

Vol 29 (18) ◽

pp. 4891-4905 ◽

Cited By ~ 16

Author(s):

Santhi Pondugula ◽

Daniel W. Neef ◽

Warren P. Voth ◽

Russell P. Darst ◽

Archana Dhasarathy ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Cell Cycle Progression ◽

Mitotic Cell ◽

Dependent Manner ◽

Phosphate Homeostasis ◽

Periodic Cell ◽

M Phase ◽

Cycle Dependent ◽

Nutrient Homeostasis

ABSTRACT Cells devote considerable resources to nutrient homeostasis, involving nutrient surveillance, acquisition, and storage at physiologically relevant concentrations. Many Saccharomyces cerevisiae transcripts coding for proteins with nutrient uptake functions exhibit peak periodic accumulation during M phase, indicating that an important aspect of nutrient homeostasis involves transcriptional regulation. Inorganic phosphate is a central macronutrient that we have previously shown oscillates inversely with mitotic activation of PHO5. The mechanism of this periodic cell cycle expression remains unknown. To date, only two sequence-specific activators, Pho4 and Pho2, were known to induce PHO5 transcription. We provide here evidence that Mcm1, a MADS-box protein, is essential for PHO5 mitotic activation. In addition, we found that cells simultaneously lacking the forkhead proteins, Fkh1 and Fkh2, exhibited a 2.5-fold decrease in PHO5 expression. The Mcm1-Fkh2 complex, first shown to transactivate genes within the CLB2 cluster that drive G2/M progression, also associated directly at the PHO5 promoter in a cell cycle-dependent manner in chromatin immunoprecipitation assays. Sds3, a component specific to the Rpd3L histone deacetylase complex, was also recruited to PHO5 in G1. These findings provide (i) further mechanistic insight into PHO5 mitotic activation, (ii) demonstrate that Mcm1-Fkh2 can function combinatorially with other activators to yield late M/G1 induction, and (iii) couple the mitotic cell cycle progression machinery to cellular phosphate homeostasis.

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