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Related Experiment Videos

Decrease in glycolytic flux in Saccharomyces cerevisiae cdc35-1 cells at restrictive temperature correlates with a

L J Oehlen1, M E Scholte, W de Koning

  • 1E. C. Slater Institute, BioCentrum, University of Amsterdam, The Netherlands.

Microbiology (Reading, England)
|August 1, 1994
PubMed
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In Saccharomyces cerevisiae, a temperature shift increased cyclic adenosine monophosphate (cAMP) but decreased glycolytic flux. Glucose transport, not enzyme activity, limits glycolysis in this yeast mutant.

Area of Science:

  • Cell Biology
  • Biochemistry
  • Microbiology

Background:

  • The adenylate cyclase gene (CDC35) is crucial for regulating cyclic adenosine monophosphate (cAMP) levels in Saccharomyces cerevisiae.
  • Thermosensitive mutants like cdc35-1 allow investigation of gene function under temperature stress.
  • Glycolytic flux is a key metabolic pathway determining cellular energy production.

Purpose of the Study:

  • To investigate the impact of a thermosensitive mutation in adenylate cyclase (cdc35-1) on glycolytic flux in Saccharomyces cerevisiae.
  • To determine the relationship between intracellular cAMP levels, cell cycle progression, and glycolytic activity.
  • To identify the rate-limiting step in glycolysis under restrictive temperature conditions.

Main Methods:

  • Utilized a thermosensitive Saccharomyces cerevisiae mutant (cdc35-1) and shifted it to a restrictive temperature.

Related Experiment Videos

  • Measured specific CO2 production rates and O2 consumption.
  • Estimated intracellular cAMP concentrations.
  • Assessed the specific activity of glycolytic enzymes and hexose transport rates.
  • Monitored intracellular concentrations of early glycolytic metabolites.

    • Main Results:

    • A temperature shift in the cdc35-1 mutant caused an initial increase in CO2 production, followed by a significant decrease.
    • Intracellular cAMP levels increased post-temperature shift, indicating high cAMP is insufficient for sustained glycolytic activity.
    • Glycolytic enzyme activity remained relatively constant, while hexose transport decreased.
    • Reduced hexose transport correlated with decreased glycolytic flux and lower intracellular metabolite levels.

    Conclusions:

    • High intracellular cAMP levels alone do not guarantee cell cycle progression or high glycolytic flux.
    • Glucose transport, rather than glycolytic enzyme activity, acts as a major control point for glycolytic flux in this yeast mutant.
    • A defect in hexose transport significantly impacts overall glycolytic activity under restrictive conditions.