SUMMARY: In the budding yeast Saccharomyces cerevisiae cyclic AMP (cAMP) can influence the activity of key enzymes in carbohydrate metabolism through modulation of the activity of cAMP-dependent protein kinase. One of the components involved in cAMP production is the CDC25 gene product, which can activate the RAS/adenylate cyclase pathway by promoting the exchange of guanine nucleotides bound to RAS. In two yeast strains carrying different thermosensitive alleles of the CDC25 gene, cAMP levels respond differently to an increase in growth temperature from 23 °C (permissive) to 36 °C (restrictive). In strain OL86 (cdc25-5) the estimated intracellular concentration of cAMP dropped after transfer to restrictive temperature whereas in strain ts321 (cdc25-1) the cAMP level rose under the same conditions. Despite the differences in cAMP levels the glycolytic flux in the two mutants responded in a very similar way to the shift from permissive to restrictive temperature; after the increase in the incubation temperature, the specific glycolytic flux in both cdc25-1 and cdc25-5 initially increased from about 300 nmol min-1 (mg protein)-1 to about 500 nmol min-1 (mg protein)-1 (presumably mainly as a consequence of the increase in temperature), but then gradually fell to 100-200 nmol min-1 (mg protein)-1. A similar pattern of CO2 production to that found in the two cdc25 mutants was also observed for several other thermosensitive mutants displaying a Start-II type of G1 arrest. In contrast, in a wild-type strain and in strains giving a Start-I type of G1 arrest, CO2 production did not drop after a temperature shift. The specific activities of glycolytic enzymes in the two cdc25 mutants did not show much change after the temperature shift, indicating that the decrease in glycolytic flux was not caused by a decrease in the activity of any of the glycolytic enzymes. Our data show that, at least in long-term regulation, the cAMP levels per se are not likely to be a prime factor controlling glycolytic flux.
BeckC.,
VonmeyenburgK.1968; Enzyme patterns and aerobic growth of Saccharomyces cerevisiae under various degrees of glucose limitation.. Journal of Bacteriology 96:479–486
BerntE.,
GutmannI.1974; Ethanol. Determination with alcohol dehydrogenase and NAD. . In: Methods of Enzymatic Analysis3 pp. 1499–1502BergmeyerH.U.
Edited by New York: Academic Press.;
BouteletF.,
PetitjeanA.,
HilgerF.1985; Yeast cdc35 mutants are defective in adenylate cyclase and are allelic with cyrl mutants while CAS1 a new gene, is involved in the regulation of adenylate cyclase.. EMBO Journal 4:2635–2641
CamonisJ.H.,
KalekineM.,
GondreB.,
GarreauH.,
Boy-MarcotteE.,
JacquetM.1986; Characterization, cloning and sequence analysis of the CDC25 gene which controls the cyclic AMP level of Saccharomyces cerevisiae.
. EMBO Journal 5:375–380
CliftonD.,
FraenkelD.G.1983; Fructose 2,6-bisphosphate and fructose 6-phosphate 2-kinase in Saccharomyces cerevisiae in relation to metabolic state in wild type and fructose 6-phosphate 1-kinase mutant strain.. Journal of Biological Chemistry 258:9245–9249
De KoningW.,
GroeneveldK.,
OehlenL.J.W.M.,
BerdenJ.A.,
VandamK.
1991; Changes in the activities of key enzymes of glycolysis during the cell cycle in yeast - a rectification.. Journal of Genera! Microbiology 137:971–976
De KoningW.,
VandamK.1992; A method for the determination of changes of glycolytic metabolites in yeast on a subsecond time scale using extraction at neutral pH.. Analytical Biochemistry 204:118–123
FraenkelD.G.1982; Carbohydrate metabolism.. In The Molecular Biology of the Yeast Saccharomyces pp. 1–37BroachJ.R.
Edited by Cold Spring Harbor NY: Cold Spring Harbor Laboratory.;
FrancoisJ.,
Van SchaftingenE.,
HersH.G.1984; The mechanism by which glucose increases fructose 2,6-bisphosphate concentration in Saccharomyces cerevisiae.
. European Journal of Biochemistry 145:187–193
GarreauH.,
CamonisJ.H.,
GuittonC.,
JacquetM.1990; The Saccharomyces cerevisiae CDC25 gene product is a 180 kDa polypeptide and is associated with a membrane fraction.. FEES Letters 269:53–59
HersH.G.,
FrancoisJ.,
Van SCHAFTINGENE.1985; Fructose- 2, 6-bisphosphate versus cyclic AMP in the liver and in lower eukaryotic cells.. Current Topics in Cellular Regulation 27:399–410
JonesS.,
VignaisM.L.,
BroachJ.R.1991; The CDC25 protein of Saccharomyces cerevisiae promotes exchange of guanine nucleotides bound to Ras.. Molecular and Cellular Biology 11:2641–2646
LagunasR.,
DominguezC.
1982; Mechanisms of appearance of the Pasteur effect in Saccharomyces cerevisiae: inactivation of sugar transport system.. Journal of Bacteriology 152:19–25
MarteganiE.,
VanoniM.,
BaroniM.1984; Macromolecular syntheses in the cell cycle mutant cdc25 of budding yeast.. European Journal of Biochemistry 144:205–210
NisslerK.,
OttoA.,
SchellenbergerW.,
HofmannE.1983; Similarity of activation of yeast phosphofructokinase by AMP and fructose-2,6-bisphosphate.. Biochemical and Biophysical Research Communications 111:294–300
NovakB.,
MitchisonJ.M.1986; Change in the rate of CO2 production in synchronous cultures of the fission yeast Schizo- saccharomyces pombe : a periodic cell cycle event that persists after the DNA-division cycle has been blocked.. Journal of Cell Science 86:191–206
PetitjeanA.,
HilgerF.,
TatchellK.1990; Comparison of thermosensitive alleles of the CDC25 gene involved in the cAMP metabolism of Saccharomyces cerevisiae.
. Genetics 124:797–806
PohligG.
1985; Phosphorylation and inactivation of yeast fructose 1,6-bisphosphatase by cyclic AMP-dependent protein kinase from yeast.. Journal of Biological Chemistry 260:13818–13823
PringleJ.R.,
HartwellL.H.1981; The Saccharomyces cerevisiae cell cycle.. In The Molecular Biology of the Yeast Saccharomyces pp. 97–142StrathernJ. N.,
BroachJ.R.
Edited by Cold Spring Harbor NY: Cold Spring Harbor Laboratory.;
PurwinC.,
LeidigF.,
HolzerH.1982; cAMP-dependent phosphorylation of fructose 1,6-bisphosphatase in yeast.. Biochemical and Biophysical Research Communications 107:1482–1489
RittenhouseJ.,
MoberlyL.,
MarcusF.1987; Phosphorylation in vivo of yeast (Saccharomyces cerevisiae) fructose-1,6- bisphosphatase at the cyclic AMP-dependent site.. Journal of Biological Chemistry 262:10114–10119
TanakaK.,
MatsumotoK.,
Toh-EA.1988; Dual regulation of the expression of the polyubiquitin gene by cyclic AMP and heat shock in yeast.. EMBO Journal 7:495–502
TortoraP.,
BirtelM.,
LenzA.G.,
HolzerH.1981; Glucose- dependent metabolic interconversion of fructose-1,6-bisphosphatase in yeast.. Biochemical and Biophysical Research Communications 100:688–695
ToveyK.C.,
OldhamK.G.,
WhelanJ.A.M.1974; A simple direct assay for cyclic AMP in plasma and other biological samples using an improved competitive binding technique.. Clinica Chimica Acta 56:221–234
UnoL,
AdachiK.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 AelstL.,
Boy-MarcotteE.,
CamonisJ.H.,
TheveleinJ.M.,
JacquetM.1990; The C-terminal part of theCDC25 gene product plays a key role in signal transduction in the glucose-induced modulation of cAMP level inSaccharomyces cerevisiae.
. European Journal of Biochemistry 193:675–680
Van AelstL.,
JansA.W.H.,
TheveleinJ.M.1991; Involvement of thecdc25 gene product in the signal transmission pathway of the glucose-induced RAS-mediated cAMP signal in the yeastSaccharomyces cerevisiae.
. Journal of General Microbiology 137:341–349
Van DoornJ.,
ValkenburgJ.A.C.,
ScholteM.E.,
OehlenL.J.W.M.,
VandrielR.,
PostmaP.W.,
NanningaN.,
VandamK.1988; Changes in activities of several enzymes involved in carbohydrate metabolism during the cell cycle.. Journal of Bac-teriology 170:4808–4815
Van SchaftingenE.,
HersH.G.1983; Fructose 2,6-bisphosphate in relation with the resumption of metabolic activity in slices of Jerusalem artichoke tubers.. FEBS Letters 164:195–200
VanoniM.,
VavassoriM.,
FrascottiG.,
MarteganiE.,
AlberghinaL.1990; Overexpression of thecdc25 gene, an upstream element of the Ras adenylyl cyclase pathway inSaccharomyces cerevisiae allows immunological identification and characterization of its gene product.. Biochemical and Biophysical Research Communications 172:61–69