Abstract
BackgroundWhen glucose is added to Saccharomyces cerevisiae grown in non-fermentable carbon sources, genes encoding ribosomal, cell-cycle, and glycolytic proteins are induced. By contrast, genes involved in mitochondrial functions, gluconeogenesis, and the utilization of other carbon sources are repressed. Glucose also causes the activation of the plasma membrane ATPase and the inactivation of gluconeogenic enzymes and mitochondrial enzymes. The goals of this study were to use the iTRAQ-labeling mass spectrometry technique to identify proteins whose relative levels change in response to glucose re-feeding and to correlate changes in protein abundance with changes in transcription and enzymatic activities. We used an experimental condition that causes the degradation of gluconeogenic enzymes when glucose starved cells are replenished with glucose. Identification of these enzymes as being down-regulated by glucose served as an internal control. Furthermore, we sought to identify new proteins that were either up-regulated or down-regulated by glucose.ResultsWe have identified new and known proteins that change their relative levels in cells that were transferred from medium containing low glucose to medium containing high glucose. Up-regulated proteins included ribosomal subunits, proteins involved in protein translation, and the plasma membrane ATPase. Down-regulated proteins included small heat shock proteins, mitochondrial proteins, glycolytic enzymes, and gluconeogenic enzymes. Ach1p is involved in acetate metabolism and is also down-regulated by glucose.ConclusionsWe have identified known proteins that have previously been reported to be regulated by glucose as well as new glucose-regulated proteins. Up-regulation of ribosomal proteins and proteins involved in translation may lead to an increase in protein synthesis and in nutrient uptake. Down-regulation of glycolytic enzymes, gluconeogenic enzymes, and mitochondrial proteins may result in changes in glycolysis, gluconeogenesis, and mitochondrial functions. These changes may be beneficial for glucose-starved cells to adapt to the addition of glucose.
Highlights
When glucose is added to Saccharomyces cerevisiae grown in non-fermentable carbon sources, genes encoding ribosomal, cell-cycle, and glycolytic proteins are induced
Down-regulation of glycolytic enzymes, gluconeogenic enzymes, and mitochondrial proteins may lead to changes in glycolysis, gluconeogenesis, and mitochondrial functions when cells are transferred from glucose-deficient medium to glucose-rich medium
Proteins identified by Isobaric Tags for Relative and Absolute Quantification (iTRAQ) To identify new proteins that are regulated by glucose, we used an experimental condition that causes the degradation of gluconeogenic enzymes during glucose refeeding
Summary
When glucose is added to Saccharomyces cerevisiae grown in non-fermentable carbon sources, genes encoding ribosomal, cell-cycle, and glycolytic proteins are induced. Genes involved in mitochondrial functions, gluconeogenesis, and the utilization of other carbon sources are repressed. We used an experimental condition that causes the degradation of gluconeogenic enzymes when glucose starved cells are replenished with glucose. Identification of these enzymes as being down-regulated by glucose served as an internal control. Glucose represses genes required for mitochondrial functions, genes encoding the gluconeogenic enzymes such as FBP1 (fructose-1,6-bisphosphatase), ICL1 (isocitrate lyase), PCK1 (phosphoenolpyruvate carboxykinase), and MLS1 (malate synthase) [4,6,17,26,28,29,30], and genes required for metabolism of sugars other than glucose and fructose (Figure 1). The Ras/PKA pathway, the Gpr/Gpa circuit, the Sch pathway, the Rgt network, and the Snf network have important roles in transcriptional regulation of glucose-repressible or glucose-inducible genes [4,6,16,25,29,34,35,36,37]
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