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Comparative transcriptome analysis of Saccharomyces cerevisiae wild type and Δasc1 strain

ABSTRACT: Asc1p and its essential orthologues in higher eukaryotes, as e.g. RACK1 in mammals, are involved in several distinct cellular signaling processes, but the implications of a total deletion have never been assessed in a comprehensive genome-wide manner. This study reveals the major cellular processes affected in a Saccharomyces cerevisiae Δasc1 deletion background via de novo proteome and transcriptome analysis, as well as subsequent phenotypical characterizations. The deletion of ASC1 reduces iron-uptake and causes nitrosative stress, both known indicators for hypoxia, which manifests in a shift of energy metabolism from respiration to fermentation in the Δasc1 strain. The impact of Asc1p in cellular metabolism is further expressed by its regulative role in the MAP kinase signal transduction pathways of invasive/filamentous growth, pheromone response and cell wall integrity. In the Δasc1 mutant strain aberrations from the expected cellular response, mediated by these pathways, can be observed and are linked to changes in protein abundances of pathway-targeted transcription factors. Evidence for the translational regulation of such transcription factors suggests that ribosomal Asc1p is involved in signal transduction pathways by controlling the biosynthesis of the respective final transcriptional regulators.

ORGANISM(S): Saccharomyces cerevisiae

PROVIDER: GSE34286 | GEO | 2012/11/30

SECONDARY ACCESSION(S): PRJNA149949

REPOSITORIES: GEO

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Comparative transcriptome analysis of Saccharomyces cerevisiae wild type and M-NM-^Tasc1 strain

Project description:Asc1p and its essential orthologues in higher eukaryotes, as e.g. RACK1 in mammals, are involved in several distinct cellular signaling processes, but the implications of a total deletion have never been assessed in a comprehensive genome-wide manner. This study reveals the major cellular processes affected in a Saccharomyces cerevisiae M-NM-^Tasc1 deletion background via de novo proteome and transcriptome analysis, as well as subsequent phenotypical characterizations. The deletion of ASC1 reduces iron-uptake and causes nitrosative stress, both known indicators for hypoxia, which manifests in a shift of energy metabolism from respiration to fermentation in the M-NM-^Tasc1 strain. The impact of Asc1p in cellular metabolism is further expressed by its regulative role in the MAP kinase signal transduction pathways of invasive/filamentous growth, pheromone response and cell wall integrity. In the M-NM-^Tasc1 mutant strain aberrations from the expected cellular response, mediated by these pathways, can be observed and are linked to changes in protein abundances of pathway-targeted transcription factors. Evidence for the translational regulation of such transcription factors suggests that ribosomal Asc1p is involved in signal transduction pathways by controlling the biosynthesis of the respective final transcriptional regulators. The transcriptional data results from from 3 biological replicates for each strain, whereas for each biological replicate 2 technical replications and dye-swaps were analyzed.

2012-11-30 | E-GEOD-34286 | biostudies-arrayexpress

The ribosomal protein Asc1/RACK1 is required for efficient translation of short mRNAs

Project description:Translation is a core cellular process carried out by a highly conserved macromolecular machine, the ribosome. There has been remarkable evolutionary adaptation of this machine through the addition of eukaryote-specific ribosomal proteins whose individual effects on ribosome function are largely unknown. Here we show that eukaryote-specific Asc1/RACK1 is required for efficient translation of mRNAs with short open reading frames that show greater than average translational efficiency in diverse eukaryotes. ASC1 mutants in S. cerevisiae display compromised translation of specific functional groups, including cytoplasmic and mitochondrial ribosomal proteins, and display cellular phenotypes consistent with their gene-specific translation defects. Asc1-sensitive mRNAs are preferentially associated with the translational ‘closed loop’ complex comprised of eIF4E, eIF4G and Pab1, and depletion of eIF4G mimics the translational defects of ASC1 mutants. Together our results reveal a role for Asc1/RACK1 in a length-dependent initiation mechanism optimized for efficient translation of genes with important housekeeping functions.

2016-04-24 | GSE61753 | GEO

Ustilago maydis rak1 mediated gene expression during exponential growth

Project description:In mammalian cells RACK1 serves as a scaffold protein that has a role in integrating inputs from different signaling pathways and affects translation through association with ribosomes. U. maydis contains a seven-WD40 repeat motif protein designated Rak1, which shows 68% identity to RACK1 and is homologous to Asc1p of Saccharomyces cerevisiae. The asc1 mutant could be complemented by introduction of U. maydis rak1. The deletion of rak1 affected cell growth, cell wall integrity and specifically attenuated cell fusion. This latter defect was caused by reduced expression of prf1 encoding the regulator for pheromone and pheromone-receptor genes. Rak1 interacts with a variety of ribosomal proteins and microarray analysis revealed that the deletion of rak1 led to severely reduced expression of rop1, an activator prf1. The constitutive expression of rop1 could rescue the defect of mfa1 expression as well as conjugation tube formation in response to pheromone induction in the rak1 mutant. Moreover, a solopathogenic rak1 mutant failed to respond to plant derived stimuli, resulting in attenuated filamentation and pathogenicity. This could be partially rescued by constitutive expression of the b heterodimer. These data suggest that rak1 is a regulator of rop1 expression with additional roles after cell fusion. Ustilago maydis strains FB1 and FB1?rak1 were grown on a rotary shaker (200 rpm) in liquid Ustilago complete medium with 1% glucose at 28°C to an OD600 of 0.5. The experiment was performed using three independent biological replicates per strain.

2011-07-05 | E-GEOD-30382 | biostudies-arrayexpress

Ustilago maydis rak1 mediated gene expression during exponential growth

2011-07-06 | GSE30382 | GEO

Multi-omics analysis of glucose-mediated signaling by a moonlighting Gβ protein Asc1/RACK1

Project description:While much is known about glucose metabolism in yeast, less is known about the receptors and signaling pathways that indicate glucose availability. We obtained metabolic profiles for wildtype and 16 mutants affecting the yeast glucose sensing pathway, comparing 0.05% glucose vs 10 min after glucose addition to 2%. First, we determined that the G protein-coupled receptor (Gpr1/Gpa2) directs early events in glucose utilization while the transceptors (Snf3/Rgt2) regulate subsequent processes and downstream products of glucose metabolism. Whereas the large G protein transmits the signal from its cognate receptor, Ras2 (but not Ras1) integrates responses from both receptor pathways. Second, we determined the relative contributions of the G protein α (Gpa2) and β (Asc1) subunits to glucose-initiated processes. We determined that Gpa2 is primarily involved in regulating carbohydrate metabolism while Asc1 is primarily involved in amino acid metabolism. Both proteins are involved in regulating purine metabolism. Collectively, our analysis reveals the molecular basis for glucose detection and the earliest events of glucose-dependent signal transduction in yeast.

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Project description:The response to bile of Lactobacillus casei BL23 and its derivative strain TC01 was investigated. TC01 strain carries a complete deletion of gene LCABL_02080 which encodes a two component signal transduction response regulator.

2012-10-10 | GSE25448 | GEO

transcriptional reponse of yeast glucose sensing pathway mutants to glucose addition

Project description:While much is known about glucose metabolism in yeast, less is known about the receptors and signaling pathways that indicate glucose availability. Here we compared wildtype and 16 mutants in yeast glucose sensing pathway for their transcriptomics profiles in 0.05% glucose vs 10 min after glucose addition to 2% glucose. With these data, we were able to define various roles of glucose sensing pathway components. We demonstrated that the G protein-coupled receptor (Gpr1/Gpa2) directed early events in glucose utilization, the transceptors (SNF3/RGT2) regulated subsequent processes and downstream products of glucose metabolism. Whereas the large G protein transmits the signal from its cognate receptor, Ras2 (but not Ras1) integrates responses from both receptor pathways. We also determined the relative contributions of the Gα (Gpa2) and Gβ (Asc1) protein subunits to glucose-initiated processes in yeast. We determined that Gpa2 is primarily involved in regulating carbohydratesugar metabolism while Asc1 is primarily involved in amino acid metabolism. Both proteins are involved in regulating purine metabolism. Collectively, our analysis reveals the molecular basis for glucose detection and the earliest events of glucose-dependent signal transduction in yeast.

2021-06-09 | GSE171846 | GEO

Quantitative Rps2 BioID to monitor Asc1-dependent rearrangement at the head region of the 40S ribosome

Project description:In a previous study we applied a quantitative proximity-labeling technique called Biotin Identification (BioID, Roux et al., 2012) to analyze the microenvironment of the Gβ-like scaffold protein Asc1 (Opitz et al., 2017). The WD40-repeat protein Asc1 binds to the head region of the small 40S ribosomal (hr40S) subunit. In this study, we analyzed the hr40S from the perspective of Rps2, a ribosomal neighbor of Asc1. We intended to confirm proteins of the Asc1 proxiOME at the hr40S and to observe changes when Asc1 was absent. References: Roux K.J., Kim D.I., Raida M., Burke B. 2012. A promiscuous biotin ligase fusion protein identifies proximal and interacting proteins in mammalian cells. J Cell Biol 196:801-810 Opitz N., Schmitt K., Hofer-Pretz V., Neumann B., Krebber H., Braus G.H., Valerius O. 2017. Capturing the Asc1p/RACK1 microenvironment at the head region of the 40S ribosome with quantitative BioID in yeast. Mol Cell Proteomics 16:2199-2218

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