Regulation of the cAMP level in the yeast Saccharomyces cerevisiae: the glucose-induced cAMP signal is not mediated by a transient drop in the intracellular pH.
Addition of glucose to derepressed cells of the yeast Saccharomyces cerevisiae is known to cause a rapid, transient increase in the cAMP level, which lasts for 1-2 min and induces a cAMP-dependent protein phosphorylation cascade. The glucose-induced cAMP signal cannot be explained solely on the basis of an increased ATP level. Transient membrane depolarization and transient intracellular acidification have been suggested as possible triggers for the cAMP peak. Addition of glucose to cells in which the plasma membrane had been depolarized still produced the increase in the cAMP level excluding membrane depolarization as the possible trigger. Using in vivo 31P NMR-spectroscopy we followed phosphate metabolism and the time course of the drop in the intracellular pH after addition of glucose with a time resolution of 15 s. Under aerobic conditions the initial pH and ATP level were high. On addition of glucose, they both showed a rapid, transient drop, which lasted for about 30 s. Under anaerobic conditions, the initial pH and ATP level were low and on addition of glucose they both increased relatively slowly compared to aerobic conditions. Several conditions were found in which the pH drop which occurs under aerobic conditions could be blocked completely without effect on the cAMP signal or without completely preventing it: addition of NH4Cl together with glucose at high extracellular pH and addition of a low concentration of glucose before a high concentration. Also, when glucose was added twice to the same cells no consistent relationship was observed between the pH drop and the cAMP peak. These results appear to exclude transient intracellular acidification as the trigger for the cAMP signal. Hence, we conclude that the effect of glucose cannot be explained on the basis of effects known to be caused by the membrane depolarizing compounds which cause increases in the cAMP level. A new, more specific kind of interaction appears to be involved.
Thevelein
J M
JM
Laboratorium voor Cellulaire Biochemie, Katholieke Universiteit te Leuven, Heverlee, Belgium.
Beullens
M
M
Honshoven
F
F
Hoebeeck
G
G
Detremerie
K
K
Griewel
B
B
den Hollander
J A
JA
Jans
A W
AW
D000242:Cyclic AMP / Q000378:metabolism*; D004140:Dinitrophenols / Q000494:pharmacology; D005947:Glucose / Q000493:pharmacokinetics / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy; D012441:Saccharomyces cerevisiae / Q000378:metabolism*
D004140:Dinitrophenols; D000242:Cyclic AMP; D005947:Glucose