Signal transduction dynamics of the protein kinase-A/phosphofructokinase-2 system in Saccharomyces cerevisiae.
This work focuses on the phosphofructokinase-2-system dynamics in Saccharomyces cerevisiae, in vivo. The investigations were dedicated to the development and implementation of appropriate theoretical and experimental methods toward evaluation of a quantitative strategy for the characterization of systemic mechanisms involved in the cAMP/protein kinase-A/phosphofructokinase-2 signal transduction cascade in yeast. Upon glucose pulse experiments, applied to glucose-limited continuous cultures of S. cerevisiae, the system response was determined with respect to alterations of intracellular metabolite concentrations or in vivo enzyme activities. Phosphofructokinase-2, in vivo, was found to be saturated with respect to both its substrates, F6P and ATP. This restriction results in an uncoupling of the enzyme activity and the signal transduction cascade from glycolytic flux, concluding that activation of phosphofructokinase-2 is exclusively a result of phosphorylation by protein kinase-A, which in turn is activated by increasing intracellular cAMP concentration after an extracellular glucose pulse. Signal processing from cAMP versus phosphofructokinase-2 also displays peculiar features implicated in a hysteresis behavior: when increasing cAMP concentration achieves a certain critical value, protein kinase-A switches into an active state. Posterior to this activation, the signal transform maintains autonomy and functional independence of further alterations of the intracellular cAMP concentration. Our observations, finally, allow the establishment of a representative model for the description of the signal transduction process via protein kinase-A in yeast.
Vaseghi
S
S
baITec-Research, Alsterkrugchaussee 374, 22335 Hamburg, Germany. vaseghi.s@baitec.de
Macherhammer
F
F
Zibek
S
S
Reuss
M
M
D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D007700:Kinetics; D008954:Models, Biological; D008962:Models, Theoretical; D010732:Phosphofructokinase-1 / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D015398:Signal Transduction; D013997:Time Factors
D000242:Cyclic AMP; D010732:Phosphofructokinase-1; D017868:Cyclic AMP-Dependent Protein Kinases; D005947:Glucose