1887

Microbiology Society logo

Volume 135, Issue 9

The Mechanism of Intracellular Acidification Induced by Glucose in Free

Abstract

Addition of glucose or fructose to cells of adapted to grow in the absence-of glucose induced an acidification of the intracellular medium. This acidification appeared to be due to the phosphorylation of the sugar since: (i) glucose analogues which are not efficiently phosphorylated did not induce internal acidification; (ii) glucose addition did not cause internal acidification in a mutant deficient in all the three sugar-phosphorylating enzymes; (iii) fructose did not affect the intracellular pH in a double mutant having only glucokinase activity; (iv) glucose was as effective as fructose in inducing the internal pH drop in a mutant deficient in phosphoglucose isomerase activity; and (v) in strains deficient in two of the three sugar-phosphorylating activities, there was a good correlation between the specific glucose-or fructose-phosphorylating activity of cell extracts and the sugar-induced internal acidification. In addition, in whole cells any of the three yeast sugar kinases were capable of mediating the internal acidification described. Glucose-induced internal acidification was observed even when yeast cells were suspended in growth medium and in cells suspended in buffer containing K, which supports the possible signalling function of the glucose-induced internal acidification. Evaluation of internal pH by following fluorescence changes of fluorescein-loaded cells indicated that the change in intracellular pH occurred immediately after addition of sugar. The apparent for glucose in this process was 2 m. Changes in both the internal and external pH were determined and it was found that the internal acidification induced by glucose was followed by a partial alkalinization coincident with the initiation of H efflux. This reversal of acidification could be due to the activity of the H-ATPase, since it was inhibited by diethylstilboestrol. Coincidence between internal alkalinization and the H efflux was also observed after addition of ethanol.

  • Received:
  • Accepted:
  • Published Online:
© Society for General Microbiology, 1989
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-135-9-2413
1989-09-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/135/9/mic-135-9-2413.html?itemId=/content/journal/micro/10.1099/00221287-135-9-2413&mimeType=html&fmt=ahah

References

  1. Aguilera A. 1986; Deletion of the phosphoglucose isomerase structural gene makes growth and sporula-tion glucose dependent in Saccharomyces cerevisiae . Molecular and General Genetics 204:310–316
     [Google Scholar]
  2. Ballou C.E. 1982; Yeast cell wall and cell surfaces. In The Molecular Biology of the Yeast Saccharomyces. Metabolism and Gene Expression pp. 335–360 Strathem N. J., Jones E. W., Broach J. R. Edited by Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  3. Barnard E.A. 1975; Hexokinases from yeasts. Methods in Enzymology 42C:6–20
     [Google Scholar]
  4. Beullens M., Mbonyi K., Geerts L., Gladines D., Detremerie K., Jans A.W.H., Thevelein J.M. 1988; Studies on the mechanism of the glucose-induced cAMP signal in glycolysis and glucose repression mutants of Saccharomyces cerevisiae . European Journal of Biochemistry 172:227–231
     [Google Scholar]
  5. Bisson L.F., Fraenkel G. 1983; Involvement of kinases in glucose and fructose uptake by Saccharomyces cerevisiae . Proceedings of the National Academy of Sciences of the United States of America 80:1730–1734
     [Google Scholar]
  6. Blatt M.R., Slayman C.L. 1987; Role of “active” postassium transport in the regulation of cytoplasmic pH by nonanimal cells. Proceedings of the National Academy of Sciences of the United States of America 84:2737–2741
     [Google Scholar]
  7. Borst-Pauwels G.W.F.H. 1981; Ion transport in yeast. Biochimica et biophysica acta 650:88–127
     [Google Scholar]
  8. Busa W.B., Nuccitelli R. 1984; Metabolic regulation via intracellular pH. American Journal of Physiology 246:409–438
     [Google Scholar]
  9. Caspani G., Tortora P., Hanozet G.M., Guerritore A. 1985; Glucose-stimulated cAMP increase may be mediated by intracellular acidification in Saccharomyces cerevisiae . FEBS Letters 186:75–79
     [Google Scholar]
  10. Duro A.F., Serrano R. 1981; Inhibition of succinate production during yeast fermentation by deenergization of the plasma membrane. Current Microbiology 6:111–113
     [Google Scholar]
  11. Entian K.D., Mecke D. 1981; Genetic evidence for a role of hexokinase isozyme PII in carbon catabolite repression in Saccharomyces cerevisiae . Journal of Biological Chemistry 257:870–874
     [Google Scholar]
  12. Eraso P., Mazon M.J., Gancedo J.M. 1987; Internal acidification and cAMP increase are not correlated in Saccharomyces cerevisiae . European Journal of Biochemistry 165:671–674
     [Google Scholar]
  13. Fernández R., Herrero P., Moreno F. 1985; Inhibition and inactivation of glucose-phosphorylat-ing enzymes from Saccharomyces cerevisiae by d-xylose. Journal of General Microbiology 131:2705–2709
     [Google Scholar]
  14. Fraenkel D.G. 1982; Carbohydrate metabolism. In The Molecular Biology of the Yeast Saccharomyces. Metabolism and Gene Expression pp. 1–37 Strathem N. J., Jones E. W., Broach J. R. Edited by Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  15. Gancedo C., Gancedo J.M. 1985; Phosphorylation of 3-O-methyl-d-glucose and catabolite repression in yeast. European Journal of Biochemistry 148:593–597
     [Google Scholar]
  16. Gancedo J.M., Clifton D., Fraenkel D.G. 1977; Yeast hexokinase mutants. Journal of Biological Chemistry 252:4443–4444
     [Google Scholar]
  17. Goffeau A., Slayman C.W. 1981; The proton-translocating ATPase of the fungal plasma membrane. Biochimica et biophysica acta 639:197–223
     [Google Scholar]
  18. Grinstein S., Rothstein A. 1986; Mechanisms of regulation of the Na+/H+ exchanger. Journal of Membrane Biology 90:1–12
     [Google Scholar]
  19. Grinstein S., Goetz-Smith J.D., Cohen S. 1987; Cytoplasmic free Ca2+ and the intracellular pH of lymphocytes. In Cell Calcium and the Control of Membrane Transport Society for General Physiology Series, vol.42 pp. 215–228 Mandel L. J., Eaton D. C. Edited by New York: Rockefeller University Press;
     [Google Scholar]
  20. Henderson L.M., Chappell J.B., Jones O.T.G. 1988; Internal pH changes associated with the activity of NADPH oxidase of human neutrophils. Biochemical Journal 251:563–567
     [Google Scholar]
  21. Henry S.A. 1982; Membrane lipids of yeast: Biochemical and genetic studies. In The Molecular Biology of the Yeast Saccharomyces. Metabolism and Gene Expression pp. 101–158 Strathem N. J., Jones E. W., Broach J. R. Edited by Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  22. Den Hollander J.A., Ugurbil K., Brown T.R., Shulman R.G. 1981; Phosphorus-31 nuclear magnetic resonance studies of the effect of oxygen upon glycolysis in yeast. Biochemistry 20:5871–5880
     [Google Scholar]
  23. Ives H.E., Daniel T.O. 1987; Interrelationship between growth factor-induced pH changes and intracellular Ca2+ . Proceedings of the National Academy of Sciences of the United States of America 84:1950–1954
     [Google Scholar]
  24. Madshus I.H. 1988; Regulation of internal pH in eucaryotic cells. Biochemical Journal 250:1–8
     [Google Scholar]
  25. Mazón M.J., Gancedo J.M., Gancedo C. 1982; Phosphorylation and inactivation of yeast fructose-bisphosphatase in vivo by glucose and proton ionophores. European Journal of Biochemistry 127:605–608
     [Google Scholar]
  26. Mbonyi K., Beullens M., Detremerie K., Geerts L., Thevelein J.M. 1988; Requirement of one functional RAS gene and inability of an oncogenic ras variant to mediate the glucose-induced cyclic AMP signal in the yeast Saccharomyces cerevisiae . Molecular and Cellular Biology 8:3051–3057
     [Google Scholar]
  27. Nicolay K., Scheffers W.A., Bruinenberg P.M., Kaptein R. 1982; Phosphorus-31 nuclear magnetic resonance studies of intracellular pH. Phosphate compartmentation and phosphate transport in yeast. Archives of Microbiology 133:83–89
     [Google Scholar]
  28. Ongjoco R., Szkutnicka K., Cirillo V.P. 1987; Glucose transport in vesicles reconstituted from Saccharomyces cerevisiae membranes and liposomes. Journal of Bacteriology 169:2926–2931
     [Google Scholar]
  29. De La Peña P., Barros F., Gascon S., Ramos S., Lazo P.S. 1982; The electrochemical proton gradient of Saccharomyces. Role of potassium. European Journal of Biochemistry 123:447–453
     [Google Scholar]
  30. Van Der Plaat J.B., Van Soligen P. 1974; Cyclic 3´,5´ adenosine monophosphate stimulates threalase degradation in baker’s yeast. Biochemical and Biophysical Research Communications 56:580–587
     [Google Scholar]
  31. Purich D.L., Fromm H.J., Rudolph F.B. 1973; The hexokinases: kinetic, physical and regulatory properties. Advances in Enzymology 39:249–326
     [Google Scholar]
  32. Purwin C., Nicolay K., Scheffers A., Holzer H. 1986; Mechanism of control of adenylate cyclase activity in yeast by fermentable sugars and carbonyl cyanide m-chlorophenylhydrazone. Journal of Biological Chemistry 261:8744–8749
     [Google Scholar]
  33. Serrano R. 1980; Effect of ATPase inhibitors on the proton pump of respiratory-deficient yeast. European Journal of Biochemistry 105:419–424
     [Google Scholar]
  34. Serrano R. 1984; Plasma membrane ATPase of fungi and plants as a novel type of proton pump. Current Topics on Cellular Regulation 23:87–126
     [Google Scholar]
  35. Siverio J.M., Valdes-Hevia M.D., Gancedo C. 1986; Toxicity of 3-O-methylglucose in yeast is due to its phosphorylation by glucokinase. FEBS tetters 194:39–42
     [Google Scholar]
  36. Slavik J. 1982; Intracellular pH of yeast measured with fluorescent probes. FEBS tetters 140:22–26
     [Google Scholar]
  37. Sols A., De La Fuente G., Villar-Palasi C., Asensio C. 1958; substrate specificity of and some other properties of baker’s yeast hexokinase. Biochimica et biophysica acta 30:92–101
     [Google Scholar]
  38. Thevelein J.M., Beullens M., Honshoven F., Hoebeeck G., Detremerie K., Griewel B., Den Hollander J., Jans A.W.H. 1987a; Regulation of the cAMP level in the yeast Saccharomyces cerevisiae: intracellular pH and the effect of membrane depolarizing compounds. Journal of General Microbiology 133:2191–2196
     [Google Scholar]
  39. Thevelein J.M., Beullens M., Honshoven F., Hoebeeck G., Detremerie K., Griewel B., Den Hollander J., Jans A.W.H. 1987b; 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. Journal of General Microbiology 133:2197–2205
     [Google Scholar]
  40. Valle E., Bergillos L., Gascon S., Parra F., Ramos S. 1986; Trehalase activation in yeast is mediated by internal acidification. European Journal of Biochemistry 154:247–251
     [Google Scholar]
  41. Valle E., Bergillos L., Ramos S. 1987; External K+ affects the internal acidification caused by the addition of glucose to yeast cells. Journal of General Microbiology 133:535–538
     [Google Scholar]
  42. Wurst M., Sigler K., Knotkova A. 1980; Gas chromatographic determination of extracellular metabolites produced by baker’s yeast during glucose-induced acidification. Folia microbiologica 25:306–310
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-135-9-2413
Loading
The Mechanism of Intracellular Acidification Induced by Glucose in Saccharomyces cerevisiae
Microbiology 135, 2413 (1989); https://doi.org/10.1099/00221287-135-9-2413
/content/journal/micro/10.1099/00221287-135-9-2413
/content/journal/micro/10.1099/00221287-135-9-2413
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error