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

ABSTRACT: 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.

ORGANISM(S): Yeast Saccharomyces Cerevisiae

TISSUE(S): Yeast Cells

SUBMITTER: Susan Sumner

PROVIDER: ST001786 | MetabolomicsWorkbench | Mon May 10 00:00:00 BST 2021

REPOSITORIES: MetabolomicsWorkbench

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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.

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A genome-wide analysis of carbon catabolite repression in Schizosaccharomyces pombe [RNA-seq]

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A genome-wide analysis of carbon catabolite repression in Schizosaccharomyces pombe [ChIP-seq]

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Transcription profiling and proteomic profiling of Saccharomyces cerevisiae wild-type and GPA2 mutant strains

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Project description:Expression of the yeast Cth2 protein stimulates degradation of mRNAs encoding proteins with Fe-dependent functions in metabolism, in iron storage and in other cellular processes. We demonstrate that in response to Fe deprivation, the Cth2-homologue, Cth1, stimulates specific degradation of mRNAs involved in mitochondrially localized activities that include respiration and amino acid biosynthesis. Furthermore, yeast cells grown under Fe deprivation accumulate mRNAs encoding proteins that function in glucose metabolism. These studies demonstrate a reprogramming of cellular metabolism during Fe-starvation dependent on the coordinated activities of two mRNA binding proteins. Experiment Overall Design: cth1cth2 cells independently transformed with pRS416 (V), pRS416-CTH1 (C) or pRS416-CTH2 (T) were grown in triplicate in SC-Ura containing 100 Î¼M BPS until exponential growth phase (approximately 6hrs) at 30 degrees, RNA was extracted using a standard glass beads protocol, labeled and hybridized to Yeast Genome S98 Afffymetrix arrays.

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