Measurements of intracellularATP provide new insight into the regulation of glycolysis in the yeast Saccharomyces cerevisiae

Ytting, Cecilie K.; Fuglsang, Anja Thoe; Hiltunen, J. Kalervo; Kastaniotis, Alexander J.; Özalp, Veli Cengiz; Nielsen, Lise Junker; Olsen, Lars Folke

doi:10.1039/c1ib00108f

Cited by 27 publications

(40 citation statements)

References 35 publications

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“…First, it was observed that the mitochondrial membrane potential (Δψ m ) oscillates with the same frequency as NADH. Next, it was found that mitochondrial F 1 F 0 ATPase activity seems to be important for the oscillations to take place . This experimental evidence for the participation of ATPase activity in the regulation of glycolytic oscillations is supported by studies of models of the pathway .…”

Section: Introductionmentioning

confidence: 74%

An experimental study of the regulation of glycolytic oscillations in yeast

et al. 2013

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We have studied oscillating glycolysis in the strain BY4743 and isogenic strains with deletions of genes encoding enzymes in glycolysis, mitochondrial electron transport and ATP synthesis. We found that deletion of the gene encoding the hexokinase 1 isoform does not affect the oscillations while deletion of the gene encoding the hexokinase 2 isoform results in oscillations with smaller amplitude. The latter is associated with an almost 50% decrease in hexokinase activity. Deletions in the genes encoding the a-and b-subunits of phosphofructokinase abolish the oscillations entirely. This loss in oscillatory activity is associated with a fourfold decrease in phosphofructokinase activity. Deletions of genes encoding subunits of the F 1 F 0 ATPase also inhibit the oscillations in accordance with earlier studies using for example inhibitors. Finally, we identified an apparently new control point involving the mitochondrial cytochrome c oxidase. The latter is difficult to explain as oscillatory activity entails 100% inhibition of this enzyme. The mitochondria of this strain seem to have normal F 1 F 0 ATPase activity. Overall these results support earlier experimental and model studies suggesting that in addition to processes within glycolysis also processes outside this pathway contribute to the control of the oscillatory behaviour.

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Section: Introductionmentioning

confidence: 74%

An experimental study of the regulation of glycolytic oscillations in yeast

et al. 2013

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“…However, concentrations of ATP can also be high in starved cells and start to decrease when glycolysis is induced. In the absence of glucose, ATPase activity is switched off and the intracellular ATP concentration is high (Ytting et al ., ). The accumulation of adenosine triphosphate in the cell results in inhibition of phosphofructokinase (PFK), one of the main enzymes that regulate glycolysis (Bai et al ., ).…”

Section: Resultsmentioning

confidence: 97%

Biodiversity of brewery yeast strains and their fermentative activities

Berłowska

Kręgiel

Rajkowska

2014

Yeast

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We investigated the genetic, biochemical, fermentative and physiological characteristics of brewery yeast strains and performed a hierarchical cluster analysis to evaluate their similarity. We used five different ale and lager yeast strains, originating from different European breweries and deposited at the National Collection of Yeast Cultures (UK). Ale and lager strains exhibited different genomic properties, but their assimilation profiles and pyruvate decarboxylase activities corresponded to their species classifications. The activity of another enzyme, succinate dehydrogenase, varied between different brewing strains. Our results confirmed that ATP and glycogen content, and the activity of the key metabolic enzymes succinate dehydrogenase and pyruvate decarboxylase, may be good general indicators of cell viability. However, the genetic properties, physiology and fermentation capacity of different brewery yeasts are unique to individual strains.

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“…Recently, Kloster and Olsen [62] investigated the role of intracellular ATPase activity using specific inhibitors for the three types of ATPases, namely F 0 F 1 ATPase, Pma1 and vacuolar ATPase, and showed that both inhibiting and stimulating ATPase activity suppress glycolytic oscillations. Further support of the role of ATPase in glycolytic oscillations comes from the recent study of Ytting et al (2012) [63]. However, the latter study uses a GAL1 promoter replacement pma1 mutant and does not present data for its isogenic wild type parental strain.…”

Section: Discussionmentioning

confidence: 99%

Exploring the genetic control of glycolytic oscillations in Saccharomyces Cerevisiae

et al. 2012

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BackgroundA well known example of oscillatory phenomena is the transient oscillations of glycolytic intermediates in Saccharomyces cerevisiae, their regulation being predominantly investigated by mathematical modeling. To our knowledge there has not been a genetic approach to elucidate the regulatory role of the different enzymes of the glycolytic pathway.ResultsWe report that the laboratory strain BY4743 could also be used to investigate this oscillatory phenomenon, which traditionally has been studied using S. cerevisiae X2180. This has enabled us to employ existing isogenic deletion mutants and dissect the roles of isoforms, or subunits of key glycolytic enzymes in glycolytic oscillations. We demonstrate that deletion of TDH3 but not TDH2 and TDH1 (encoding glyceraldehyde-3-phosphate dehydrogenase: GAPDH) abolishes NADH oscillations. While deletion of each of the hexokinase (HK) encoding genes (HXK1 and HXK2) leads to oscillations that are longer lasting with lower amplitude, the effect of HXK2 deletion on the duration of the oscillations is stronger than that of HXK1. Most importantly our results show that the presence of beta (Pfk2) but not that of alpha subunits (Pfk1) of the hetero-octameric enzyme phosphofructokinase (PFK) is necessary to achieve these oscillations. Furthermore, we report that the cAMP-mediated PKA pathway (via some of its components responsible for feedback down-regulation) modulates the activity of glycoytic enzymes thus affecting oscillations. Deletion of both PDE2 (encoding a high affinity cAMP-phosphodiesterase) and IRA2 (encoding a GTPase activating protein- Ras-GAP, responsible for inactivating Ras-GTP) abolished glycolytic oscillations.ConclusionsThe genetic approach to characterising the glycolytic oscillations in yeast has demonstrated differential roles of the two types of subunits of PFK, and the isoforms of GAPDH and HK. Furthermore, it has shown that PDE2 and IRA2, encoding components of the cAMP pathway responsible for negative feedback regulation of PKA, are required for glycolytic oscillations, suggesting an enticing link between these cAMP pathway components and the glycolysis pathway enzymes shown to have the greatest role in glycolytic oscillation. This study suggests that a systematic genetic approach combined with mathematical modelling can advance the study of oscillatory phenomena.

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Measurements of intracellularATP provide new insight into the regulation of glycolysis in the yeast Saccharomyces cerevisiae

Cited by 27 publications

References 35 publications

An experimental study of the regulation of glycolytic oscillations in yeast

An experimental study of the regulation of glycolytic oscillations in yeast

Biodiversity of brewery yeast strains and their fermentative activities

Exploring the genetic control of glycolytic oscillations in Saccharomyces Cerevisiae

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