Unknown,Transcriptomics,Genomics,Proteomics

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Functional and Regulatory Profiling of Energy Metabolism in Fission Yeast: dynamic gene regulation during adaptation to respiratory medium.

ABSTRACT: Fission yeast in high glucose media preferentially uses fermentation for energy production, even under aerobic conditions, an analogous metabolic programme, aerobic glycolysis, is a hallmark of cancer and enables the proliferation of tumor cells. Fission yeast, unlike budding yeast, requires mitochondrial genomes and oxidative phosphorylation for survival, mitochondrial inheritance, genome organisation and RNA processing are strikingly different between the two yeasts, and S. pombe mitochondria resemble more the human mitochondria. However, mitochondrial biology and respiratory control is poorly understood in fission yeast. In this study, we used microarrays to profile gene expression before and at six time points after the shift from fermentative to respiratory medium. Cells were grown in yeast extract based media with 3% glucose to early exponential phase (time point 0), then the carbon source was changed to 3% glycerol, 0.1% glucose. Transcript levels were monitored by microarrays at several time points after the switch (0.2, 0.5, 1, 2; 04 and 24 hours). Sample from each time point is hybrydized with sample of all pooled time points. Experiment was repeated twice with the dye swap.

INSTRUMENT(S): Axon GenePix 4000B scanning hardware

ORGANISM(S): Schizosaccharomyces pombe

SUBMITTER: JÃ¼rg BÃ¤hler

PROVIDER: E-MTAB-4518 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:When the yeast Saccharomyces cerevisiae is subjected to increasing glycolytic fluxes under aerobic conditions, there is a threshold value of the glucose uptake rate at which the metabolism shifts from being purely respiratory to mixed respiratory and fermentative. This shift is characterized by ethanol production, a phenomenon known as the Crabtree effect due to its analogy with lactate overflow in cancer cells. It is well known that at high glycolytic fluxes there is glucose repression of respiratory pathways resulting in a decrease in the respiratory capacity. Despite many years of detailed studies on this subject, it is not known whether the onset of the Crabtree effect (or overflow metabolism) is due to a limited respiratory capacity or caused by glucose-mediated repression of respiration. We addressed this issue by increasing respiration in S. cerevisiae by introducing a heterologous alternative oxidase, and observed reduced aerobic ethanol formation. In contrast, increasing non-respiratory NADH oxidation by overexpression of a water-forming NADH oxidase reduced aerobic glycerol formation. The metabolic response to elevated alternative oxidase occurred predominantly in the mitochondria, while NADH oxidase affected genes that catalyze cytosolic reactions. Moreover, NADH oxidase restored the deficiency of cytosolic NADH dehydrogenases in S. cerevisiae. These results indicate that NADH oxidase localizes in the cytosol, while alternative oxidase is directed to the mitochondria. The onset of aerobic ethanol formation is demonstrated to be a consequence of an imbalance in mitochondrial redox balancing. In addition to answering fundamental physiological questions, our findings are relevant for all biomass derived applications of S. cerevisiae. Experiment Overall Design: Heterologous gene expression in chemostats using Affymetrix Yeast Genome 2.0 arrays. Total RNA extraction and sample preparation, hybridization was done according to the manufacturer's protocol.

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Functional and Regulatory Profiling of Energy Metabolism in Fission Yeast: dynamic gene regulation during adaptation to respiratory medium.

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