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Expression profile in wild type, fkh2delta and fkh2-S2A mutants

ABSTRACT: Mitotic entry is accompanyed by the expression of a cluster of so called mitotic genes, whose activation is critical for mitosis in human and yeast cells. We found a link between the transcription machinery and cell cycle control network at mitosis in fission yeast, involving the Cdk8 kinase dependent phosphorylation of the fork head transcription factor Fkh2. We have generated a non-phosphorylatable fkh2 mutant (fkh2-S2A) also. We have used microarrays to study the gene expression changes in fkh2 deletion and phosphomutants vs Wt in exponentially growing fission yeast cells to detect how the phosphorylation may affect global and mitotic gene expression.

ORGANISM(S): Schizosaccharomyces pombe Saccharomyces cerevisiae

PROVIDER: GSE31642 | GEO | 2012/07/13

SECONDARY ACCESSION(S): PRJNA145317

REPOSITORIES: GEO

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Expression profile in wild type, fkh2delta and fkh2-S2A mutants

Project description:Mitotic entry is accompanyed by the expression of a cluster of so called mitotic genes, whose activation is critical for mitosis in human and yeast cells. We found a link between the transcription machinery and cell cycle control network at mitosis in fission yeast, involving the Cdk8 kinase dependent phosphorylation of the fork head transcription factor Fkh2. We have generated a non-phosphorylatable fkh2 mutant (fkh2-S2A) also. We have used microarrays to study the gene expression changes in fkh2 deletion and phosphomutants vs Wt in exponentially growing fission yeast cells to detect how the phosphorylation may affect global and mitotic gene expression. RNA samples were taken from exponentially growing fission yeast cells and hybridized to affymetrix microarrays: wild type, fkh2delta and fkh2-S2A mutants

2012-07-12 | E-GEOD-31642 | biostudies-arrayexpress

A new transcription factor for mitosis: In S. pombe, the RFX transcription factor Sak1 works with forkhead factors to regulate mitotic expression (ChIP-Seq)

Project description:Many genes are transcriptionally regulated as a function of the cell cycle; genes needed for mitosis are strongly cell cycle regulated. From yeasts to humans, forkhead transcription factors are involved in mitotic gene expression. However, the mechanisms of regulation are unclear, especially since forkhead transcription factors can be activators or repressors, and often function in concert with other proteins. Here, we study mitotic regulation in the fission yeast S. pombe. We find that the forkhead transcription factor Fkh2 is a repressor of many mitotic genes, while a different forkhead transcription factor, Sep1, is an activator of a subset of these genes. The activator of the majority of mitotic genes appears to be Sak1, a transcription factor of the RFX family, which has not previously been implicated in cell cycle control. 3 test samples were analyzed; 1 control (no-tag) sample was processed (each sample has 2 replicate experiments)

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Project description:The opportunistic human fungal pathogen, Candida albicans, undergoes morphological and transcriptional adaptation in the switch from commensalism to pathogenicity. Although previous gene-knockout studies have identified many factors involved in this transformation, it remains unclear how these factors are regulated to coordinate the switch. Investigating morphogenetic control by post-translational phosphorylation has generated important regulatory insights into this process, especially focusing on coordinated control by the cyclin-dependent kinase Cdc28. Here we have identified the Fkh2 transcription factor as a regulatory target of both Cdc28 and the cell wall biosynthesis kinase Cbk1, in a role distinct from its conserved function in cell cycle progression. Transcript profiling of Fkh2 deletion and non-phosphrylatable mutants in yeast and hypae revealed that the the hyphal-specific shift in the phosphorylation profile is required for the expression of genes involved in pathogenesis, host interaction and biofilm formation. We confirmed that these changes in gene expression resulted in corresponding defects in pathogenic processes

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ChIP-nexus analysis of fission yeast transcription factors Cbf11, Cbf11DBM, Cbf12, Cbf12DBM, Mga2 and Fkh2

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A new transcription factor for mitosis: In S. pombe, the RFX transcription factor Sak1 works with forkhead factors to regulate mitotic expression (array)

Project description:Many genes are transcriptionally regulated as a function of the cell cycle; genes needed for mitosis are strongly cell cycle regulated. From yeasts to humans, forkhead transcription factors are involved in mitotic gene expression. However, the mechanisms of regulation are unclear, especially since forkhead transcription factors can be activators or repressors, and often function in concert with other proteins. Here, we study mitotic regulation in the fission yeast S. pombe. We find that the forkhead transcription factor Fkh2 is a repressor of many mitotic genes, while a different forkhead transcription factor, Sep1, is an activator of a subset of these genes. The activator of the majority of mitotic genes appears to be Sak1, a transcription factor of the RFX family, which has not previously been implicated in cell cycle control. 24 samples were analyzed. Control for all experiments was the same. The samples are representative of selective spore germination in S. pombe (time course arrays for the WT as well as the mentioned mutants) or the sak1-891 ts strain restrictive temperature time course (WT and ts strain)

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Project description:Mitotic genes are one of the most strongly oscillating groups of genes in the eukaryotic cell cycle. Understanding the regulation of mitotic gene expression is a key issue in cell cycle control but is poorly understood in most organisms. Here, we find a new mitotic transcription factor, Sak1, in the fission yeast S. pombe. Sak1 belongs to the RFX family of transcription factors, which have not previously been connected to cell cycle control. Sak1 binds upstream of mitotic genes in close proximity to Fkh2, a forkhead transcription factor previously implicated in regulation of mitotic genes. We show that Sak1 is the major activator of mitotic gene expression and also confirm the role of Fkh2 as the opposing repressor. Sep1, another forkhead transcription factor, is an activator for a small subset of mitotic genes involved in septation. From yeasts to humans, forkhead transcription factors are involved in mitotic gene expression, and it will be interesting to see whether RFX transcription factors may also be involved in other organisms.

2014-08-25 | GSE60712 | GEO

A new transcription factor for mitosis: In S. pombe, the RFX transcription factor Sak1 works with forkhead factors to regulate mitotic expression (array)

2014-08-25 | GSE60718 | GEO

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2017-07-13 | GSE94475 | GEO

Condensin-mediated remodeling of the mitotic chromatin landscape in fission yeast

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CDK activity at the centrosome regulates the cell cycle

Project description:Cyclin-dependent kinases (CDKs) complexed with cyclins drive progression through the eukaryotic cell cycle. From yeast to human cells, cyclin-CDK localises to the centrosome, but the importance of this localisation is unclear. The conserved ‘hydrophobic patch’ substrate docking site on human Cyclin B1 and on the equivalent fission yeast Cdc13 mediates their localisation to the centrosome and the spindle-pole body (SPB, yeast centrosome equivalent). A hydrophobic patch mutant (HPM) of Cdc13 cannot enter mitosis, but whether this mitotic defect is due to defective SPB localisation or defective cyclin-substrate docking is unknown. Here we show that artificially restoring Cdc13HPM SPB localisation in fission yeast partially rescues both mitosis and defective CDK substrate phosphorylation at both the SPB and within the cytoplasm. In addition, we found that an HPM of the S-phase cyclin Cig2 has defective SPB localisation but is still able to perform bulk DNA synthesis. Our results demonstrate that the hydrophobic patch mediates the SPB localisation of both S- and M-phase cyclins, and that Cdc13 SPB localisation is essential for mitotic entry and for full phosphorylation of CDK substrates, supporting the view that the centrosome plays a role as a signalling hub regulating CDK cell cycle control.

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