Cyclic AMP and the Stimulation of Trehalase Activity in the Yeast Saccharomyces cerevisiae by Carbon Sources, Nitrogen Sources and Inhibitors of Protein Synthesis
Summary: Addition of glucose to acetate-grown cells of Saccharomyces cerevisiae caused a rapid transient increase in the cAMP level followed by a 10-fold, transient increase in the activity of trehalase. Exhibition bromide and acridine analogues inhibited both glucose-induced responses in a similar way, confirming the role of the cAMP signal as the second messenger in the sugar-induced activation of trehalase. When nitrogen sources or protein synthesis inhibitors were added after the transient glucose-induced increase in the trehalase activity, a rapid reactivation of trehalase occurred. In this case, however, there was only a very small increase in the cAMP level, which appeared to be insignificant. When the nitrogen source or the protein synthesis inhibitor was added together with glucose, the trehalase activity remained high for a much longer time also without a significant effect on the cAMP level. When a membrane depolarizing agent was added together with the glucose, both the trehalase activity and the cAMP level remained high. Reversibility experiments in which trehalase was activated to different degrees also showed that for high sugar-induced trehalase activation a high cAMP level is needed, while nitrogen sources stimulate trehalase activity without affecting cAMP levels. In cell extracts, both cAMP and cGMP were able to activate trehalase, the latter however only at 10-fold higher concentrations. The cGMP level in vivo was about 10-fold lower than the cAMP level and was not significantly affected by nitrogen sources or protein synthesis inhibitors. Hence, neither cAMP nor cGMP seem to be involved as the second messenger in the stimulating effect of nitrogen sources and protein synthesis inhibitors on trehalase activity in yeast. Since all other evidence obtained here and before strongly points to regulation of trehalase by a ‘cAMP-dependent’ protein kinase, we suggest that the presence of a nitrogen source in the growth medium of yeast induces the rapid synthesis of an alternative second messenger able to activate this or another protein kinase.
BartonJ. K.,
den HollanderJ. A.,
LeeT. M.,
MacLaughlinA.,
ShulmanR. G.1980; Measurement of the internal pH of yeast spores by P-31 nuclear magnetic resonance.. Proceedings of the National Academy of Sciences of the United States of America 77:2470–2473
CooperT. G.1982; Nitrogen metabolism in Saccharomyces cerevisiae
. In The Molecular Biology of the Yeast Saccharomyces. Metabolism and Gene Expression pp 39–99 Edited by
StrathernJ. N.,
JonesE. W.,
BroachJ. R.
Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
FrançoisJ.,
Van SchaftingenE.,
HersH. G.1984; The mechanism by which glucose increases fructose-2,6-bisphosphate concentration in Saccharomyces cerevisiae. A cyclic-AMP-dependent activation of phosphofructokinase 2.. European Journal of Biochemistry 145:187–193
HolzerH.1984; Mechanism and function of reversible phosphorylation of fructose-1,6-bisphosphatase in yeast.. In Molecular Aspects of Cellular Regulation vol 3 pp 143–154 Edited by
CohenP.
Amsterdam: Elsevier;
MazònM. J.,
GancedoJ. M.,
GancedoC.1982; Phosphorylation and inactivation of yeast fructose-1,6-bisphosphatase in vivo by glucose and by proton ionophores. A possible role for cAMP.. European Journal of Biochemistry 127:605–608
PurwinC.,
LeidigF.,
HolzerH.1982; Cyclic AMP-dependent phosphorylation of fructose-1,6-bisphosphatase in yeast.. Biochemical and Biophysical Research Communications 107:1482–1489
RousseauP.,
HalvorsonH. O.1973; Macromolecular synthesis during the germination of Saccharomyces cerevisiae spores.. Journal of Bacteriology 113:1289–1295
TheveleinJ. M.,
JonesK. -A.1983; Reversibility characteristics of glucose-induced trehalase activation associated with the breaking of dormancy in yeast ascospores.. European Journal of Biochemistry 136:583–587
TheveleinJ. M.,
den HollanderJ. A.,
ShulmanR. G.1982; Changes in the activity and properties of trehalase during early germination of yeast ascospores: correlation with trehalose breakdown as studied by in vivo13C NMR.. Proceedings of the National Academy of Sciences of the United States of America 79:3503–3507
TheveleinJ. M.,
Van LaereA. J.,
BeullensM.,
Van AsscheJ. A.,
CarlierA. R.1983; Glucose-induced trehalase activation and trehalose mobilization during early germination of Phycomyces blakesleeanus spores.. Journal of General Microbiology 129:719–726
TortoraP.,
BurliniN.,
HanozetG. M.,
GuerritoreA.1982; Effect of caffeine on glucose-induced inactivation of gluconeogenetic enzymes in Saccharomyces cerevisiae. A possible role of cyclic AMP.. European Journal of Biochemistry 126:617–622
TrevillyanJ. M.,
PallM. L.1979; Control of cyclic adenosine 3ʹ,5ʹ-monophosphate levels by depolarizing agents in fungi.. Journal of Bacteriology 138:397–403
UnoI.,
MatsumotoK.,
AdachiK.,
IshikawaT.1983; Genetic and biochemical evidence that trehalase is a substrate of cAMP-dependent protein kinase in yeast.. Journal of Biological Chemistry 258:10867–10872
Van der PlaatJ. B.1974; Cyclic 3ʹ,5ʹ-adenosine monophosphate stimulates trehalose degradation in baker's yeast.. Biochemical and Biophysical Research Communications 56:580–587
Van SolingenP.,
Van der PlaatJ. B.1975; Partial purification of the protein system controlling the breakdown of trehalose in baker's yeast.. Biochemical and Biophysical Research Communications 62:553–560
WiemkenA.,
SchellenbergM.1982; Does a cyclic AMP-dependent phosphorylation initiate the transfer of trehalase from the cytosol into the vacuoles in Saccharomyces cerevisiae?
. FEBS Letters 150:329–331
Cyclic AMP and the Stimulation of Trehalase Activity in the Yeast Saccharomyces cerevisiae by Carbon Sources, Nitrogen Sources and Inhibitors of Protein Synthesis