Identification of phosphorylation sites and binding proteins of Saccharomyces cerevisiae Tda1
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ABSTRACT: Tda1 in Saccharomyces cerevisiae is a protein kinase that is activated in response to glucose starvation. However, how it is activated has not been investigated for a long time. In this analysis, yeast cells were grown in high (2%) or low (0.05%) glucose medium, and the lysate were subjected to immunoprecipitation of Tda1-3xHA with an anti HA antibody. Proteins in the immnoprecipitated fractions were digested and peptides were analyzed by MS/MS.
Project description:Saccharomyces cerevisiae CEN.PK113-1A was grown in glucose-limited chemostat culture with 0%, 0.5%, 1.0%, 2.8% or 20.9% O2 in the inlet gas (D= 0.10 /h, pH5, 30C).
Project description:In response to limited nitrogen and abundant carbon sources, diploid Saccharomyces cerevisiae strains undergo a filamentous transition in cell growth as part of pseudohyphal differentiation. Use of the disaccharide maltose as the principal carbon source, in contrast to the preferred nutrient monosaccharide glucose, has been shown to induce a hyper-filamentous growth phenotype in a strain deficient for GPA2 which codes for a Galpha protein component that interacts with the glucose-sensing receptor Gpr1p to regulate filamentous growth. In this report, we compare the global transcript and proteomic profiles of wild-type and Gpa2p deficient diploid yeast strains grown on both rich and nitrogen starved maltose media. We find that deletion of GPA2 results in significantly different transcript and protein profiles when switching from rich to nitrogen starvation media. The results are discussed with a focus on the genes associated with carbon utilization, or regulation thereof, and a model for the contribution of carbon sensing/metabolism-based signal transduction to pseudohyphal differentiation is proposed. Keywords: Saccharomyces cerevisiae, nitrogen starvation, maltose, pseudohyphal differentiation, yeast, expression profiling
Project description:We propose a carbon source dependent genetic regulatory network for the budding yeast Saccharomyces cerevisiae, derived from quantitative proteomic analyses integrated with bioinformatics knowledge of regulatory pathways and protein interactions. The proposed network, comprising 1247 transcription factor interactions and 126 chaperone interactions, defines the proteome shift in the cell when growing under different carbon sources. We used a label-free proteomics strategy to quantify alterations in protein abundance for S. cerevisiae when grown on minimal media using glucose, galactose, maltose and trehalose as sole carbon sources.
Project description:In this study we investigated the transcriptional response of the yeast Saccharomyces cerevisiae to potassium starvation. To this end yeast cells were grown for 60 min in media without potassium or in media with a standard potassium concnetration (50 mM KCl). Using Serial Analysis of Gene Expression (SAGE)-tag sequencing the effect of potassium starvation on the transcriptome was determined.
Project description:In the present study transcriptome and proteome of recombinant, xylose-utilising S. cerevisiae grown in aerobic batch cultures on xylose were compared with glucose-grown cells both in glucose repressed and derepressed states. The aim was to study at genome-wide level how signalling and carbon catabolite repression differed in cells grown on either glucose or xylose. The more detailed knowledge about is xylose sensed as a fermentable carbon source, capable of catabolite repression like glucose, or is it rather recognised as a non-fermentable carbon source is important in achieving understanding for further engineering this yeast for more efficient anaerobic fermentation of xylose.
Project description:The energy metabolism pathways are significantly influenced by the available carbon sources. In Saccharomyces cerevisiae, energy is primarily produced through aerobic respiration during glycerol cultivation, a process believed to depend on the yeast AMPK (AMP-activated protein kinase) homolog, Snf1. It has been reported that Snf1 increases the expression of respiratory genes by phosphorylating transcriptional activators in environments where glucose is unavailable. We discovered that Tda1, activated by Snf1, phosphorylates Hxk2. Hxk2 has been reported to function as a transcriptional repressor. Therefore, we analyzed how Tda1 affects the expression of respiratory genes by RNA-seq analysis of glycerol-cultured cells.
Project description:This time course microarray experiment was performed on Saccharomyces cerevisiae to determine the global gene expression alterations due to 3-trifluoromethyl-4-nitrophenol (TFM) exposure over time. In this experiment, yeast grown in standard, glucose-containing media were treated with 0.05mM TFM over a four hour period.
Project description:Characterization of Saccharomyces cerevisiae whole cell proteome comparing WT and PMR1 knockout cells grown for 24 h on synthetic minimal medium containing glucose as carbon source (shaking at 140 rpm, 28ºC)
Project description:Saccharomyces cerevisiae cannot metabolize cellobiose in nature. Here, S. cerevisiae was engineered to achieve cellobiose utilization by introducing both a cellodextrin transporter gene (cdt-1) and an intracellular β-glucosidase gene (gh1-1) from Neurospora crassa. We sequenced mRNA from anaerobic exponential cultures of engineered S. cerevisiae grown on cellobiose or glucose as a single carbon source in biological triplicate. Differences in gene expression between cellobiose and glucose metabolism revealed by RNA deep sequencing indicated that cellobiose metabolism induced mitochondrial activation and reduced amino acid biosynthesis under fermentation conditions.