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A PP2A-Pab1-mediated crosstalk between TORC1 & TORC2 regulates the differentiation response in fission yeast

ABSTRACT: A PP2A-Pab1-mediated crosstalk between TORC1 & TORC2 regulates the differentiation response in fission yeast

PROVIDER: PRJEB11998 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR1162930.fastq.gz	Fastqsanger.gz
	ERR1162940.fastq.gz	Fastqsanger.gz
	ERR1162931.fastq.gz	Fastqsanger.gz
	ERR1162941.fastq.gz	Fastqsanger.gz
	ERR1162932.fastq.gz	Fastqsanger.gz

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0549_2 Fission yeast Pab1 interactome

Project description:We performed mass-spectrometry analysis of proteins co-purifying with the Pab1, the B55 regulatory subunit of the PP2A protein phosphatase complex. Pab1 was tagged with YFP at the C-terminus and expressed from its endogenous promoter at its chromosomal locus. After one hour of nitrogen starvation, Pab1-YFP was pulled down and co-purifying proteins were analysed by mass-spectrometry. Our results revealed the presence of all components of PP2A protein complexes including Paa1, the catalytic subunits Ppa1, Ppa2, and Ppa3.

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A PP2A-Pab1-mediated crosstalk between TORC1 & TORC2 regulates the differentiation response in fission yeast

Project description:RNA-sequencing of S. pombe wild type, ppa1 mutants, and tor1 mutants, grown under condition of normal nitrogen and under nitrogen starvation.

2016-10-31 | E-MTAB-4106 | biostudies-arrayexpress

E-MTAB-4106

Project description:A PP2A-Pab1-mediated crosstalk between TORC1 & TORC2 regulates the differentiation response in fission yeast

2016-11-09 | E-MTAB-4106 | ExpressionAtlas

Fission Yeast TORC1 Promotes Cell Proliferation through Sfp1, a Transcription Factor Involved in Ribosome Biogenesis

Project description:To study the TORC1-mediated transcriptional regulation of the genes involved in ribosome biosynthesis in fission yeast

2023-04-01 | GSE201580 | GEO

State Transitions in the TORC1 Signaling Pathway and Information Processing in S. cerevisiae

Project description:TOR kinase complex I (TORC1) is a key regulator of cell growth and metabolism in all eukaryotes. Previous studies in yeast have shown that three GTPases, Gtr1, Gtr2 and Rho1, bind TORC1 in nitrogen and amino acid starvation conditions to block phosphorylation of the S6 kinase Sch9 and activate protein phosphatase 2A (PP2A). This leads to down-regulation of 450 Sch9-dependent protein and ribosome synthesis genes, and up-regulation of 100 PP2A-dependent nitrogen assimilation and amino acid synthesis genes. Here, using bandshift assays and microarray measurements, we show that the TORC1 pathway also populates three other stress/starvation states. First, in glucose starvation conditions, the AMP activated protein kinase (AMPK/Snf1) and at least one other factor, push the TORC1 pathway into an off state where Sch9- branch signaling and PP2A-branch signaling are both inhibited. Remarkably, the TORC1 pathway remains in the glucose starvation (PP2A off) state even when cells are simultaneously starved for nitrogen and glucose. Second, in osmotic stress, the MAPK Hog1/p38 drives the TORC1 pathway into a different Sch9 off, PP2A off state, where PP2A-branch signaling can still be activated by nitrogen starvation. Third, in oxidative stress and heat stress, TORC1-Sch9 signaling is blocked while weak PP2A-branch signaling occurs. Together, our data show that the TORC1 pathway acts an information-processing hub, activating different genes in different conditions to ensure that available energy is allocated to drive growth, amino acid synthesis or a stress response, depending on the needs of the cell.

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TORC1 signaling pathway inhibition by rapamycin and caffeine affect lifespan, global gene expression and cell proliferation of fission yeast

Project description:Target of rapamycin complex 1 (TORC1) is implicated in growth control and aging from yeast to humans. Fission yeast is emerging as a popular model organism to study TOR signaling, although rapamycin has been thought to not affect cell growth in this organism. Here we analyzed the effects of rapamycin and caffeine, singly and combined, on multiple cellular processes in fission yeast. The two drugs led to diverse and specific phenotypes that depended on TORC1 signaling pathway inhibition, including prolonged chronological lifespan, inhibition of global translation, inhibition of cell growth and division, and reprogramming of global gene expression mimicking nitrogen starvation. Rapamycin and caffeine differentially affected these various TORC1-dependent processes. Combined drug treatment augmented most phenotypes and effectively blocked cell growth. Although rapamycin showed a much more subtle effect on global translation than did caffeine, rapamycin was more effective in prolonging chronological lifespan. Rapamycin prolonged the lifespan of non-growing cells only when applied during the growth phase but not when applied after cells had stopped proliferation. The doses of rapamycin and caffeine strongly correlated with growth inhibition and with lifespan extension. This comprehensive analysis will inform future studies into TORC1 function and cellular aging in fission yeast and beyond.

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Project description:Extracellular cues regulate cell fate, and this is mainly achieved through the engagement of specific transcriptional programs. The TORC1 and TORC2 complexes mediate the integration of nutritional cues to cellular behavior, but their interplay is poorly understood. Here, we use fission yeast to investigate how phosphatase activity participates in this interplay during the switch from proliferation to sexual differentiation. We find that loss of PP2A-B55Pab1 enhances the expression of differentiation-specific genes and leads to premature conjugation. pab1 deletion brings about a transcriptional profile similar to TORC1 inactivation, and deletion of pab1 overcomes the repression of differentiation genes in cells overexpressing TORC1. Importantly, we show that this effect is mediated by an increased TORC2-AKT (Gad8) signaling. Under nutrient-rich conditions, PP2A-B55Pab1 dephosphorylates Gad8 Ser546, repressing its activity. Conversely, TORC1 inactivation upon starvation leads to the inactivation of PP2A-B55Pab1 through the Greatwall-Endosulfin pathway. This results in the activation of Gad8 and the commitment to differentiation. Thus, PP2A-B55Pab1 enables a crosstalk between the two TOR complexes that controls cell-fate decisions in response to nutrient availability.

| S-EPMC5266790 | biostudies-other

Multilayered control of protein turnover by TORC1 and ATG1

Project description:To study TORC1-Atg1 signaling comprehensively and to identify potential additional foci of crosstalk, we chose a mass spectrometry (MS)-based phosphoproteomics strategy combing global proteomics screens in vivo with targeted analyses using in vitro kinase reactions. We present the currently largest compendium of rapamycin-sensitive phosphorylation events in the yeast Saccharomyces cerevisiae, identify numerous unknown TORC1 and Atg1 downstream phosphorylation events, and characterize hitherto unknown, functionally relevant TORC1 target sites on defined Atg proteins.

2019-09-30 | PXD013271 | Pride

Fission Yeast TORC1 Promotes Cell Proliferation through Sfp1, a Transcription Factor Involved in Ribosome Biogenesis

Project description:Fission Yeast TORC1 Promotes Cell Proliferation through Sfp1, a Transcription Factor Involved in Ribosome Biogenesis

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2019-07-19 | GSE133591 | GEO

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