1887

Abstract

Summary

Addition of glucose or related fermentable sugars to derepressed cells of the yeast triggers a RAS-protein-mediated cAMP signal, which induces a protein phosphorylation cascade. Yeast strains without a functional gene were deficient in basal cAMP synthesis and in the glucose-induced cAMP signal. Addition of dinitrophenol, which in wild-type strains strongly stimulates cAMP synthesis by lowering intracellular pH, did not enhance the cAMP level. disruption mutants, in which the basal cAMP level was restored by the oncogene or by disruption of the gene () coding for the high-affinity phosphodiesterase, were still deficient in the glucose- and acidification-induced cAMP responses. These results indicate that the gene product is required not only for basal cAMP synthesis in yeast but also for specific activation of cAMP synthesis by the signal transmission pathway leading from glucose to adenyl cyclase. They also show that intracellular acidification stimulates the pathway at or upstream of the CDC25 protein. When shifted to the restrictive temperature, cells with the temperature sensitive -5 mutation lost their cAMP content within a few minutes. After prolonged incubation at the restrictive temperature, cells with this mutation, and also those with the temperature sensitive mutation, arrested at the ‘start’ point (in G) of the cell cycle, and subsequently accumulated in the resting state G. In contrast with cells, however, the cAMP level did not decrease and normal glucose- and acidification-induced cAMP responses were observed when cells were shifted to the restrictive temperature. These results show that, in the original genetic background at least, growth arrest of -bearing cells at the restrictive temperature is not due to cAMP deficiency. Previous experiments have provided evidence for the presence of a glucose-repressible protein in the signalling pathway. Exponential-phase glucose-grown cells of a strain with overexpression of unexpectedly showed a glucose-induced cAMP signal. Control experiments, however, indicated that overexpression of caused a defect in glucose repression. Introduction of the derepression mutation in the strain with overexpression of restored glucose repression and abolished the glucose-induced cAMP signal, indicating that the CDC25 protein is not the glucose-repressible component of the signalling pathway.

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1991-02-01
2024-04-24
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References

  1. Arguelles J.-C., Mbonyi K., Van Aelst L., Vanhalewyn M., Jans A. W. H., Thevelein J. M. 1990; Absence of glucose-induced cAMP signalling in the Saccharomyces cerevisiae mutants cat 1 and cat 3 which are deficient in derepression of glucose-repressible proteins.. Archives of Microbiology 154:199–205
    [Google Scholar]
  2. Ashwell G. 1957; Colorimetric analysis of sugars. Methods in Enzymology 3:73–105
    [Google Scholar]
  3. Berridge M. J. 1987; Inositol trisphosphate and diacylglycerol: two interacting second messengers. Annual Review of Biochemistry 56:159–193
    [Google Scholar]
  4. Beullens M., Thevelein J. M. 1990; Investigation of transport-associated phosphorylation of sugar in yeast mutants (snf3) lacking high-affinity glucose transport and in a mutant (fdpl) showing deficient regulation of initial sugar metabolism.. Current Microbiology 21:39–6
    [Google Scholar]
  5. 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 the yeast Saccharomyces cerevisiae. European Journal of Biochemistry 172:227–231
    [Google Scholar]
  6. Boutelet F., Petitjean A., Hilger F. 1985; Yeast cdc35 mutants are defective in adenylate cyclase and are allelic with cyr 1 mutants while CAS1, a new gene, is involved in the regulation of adenyl cyclase.. EMBO Journal 4:2635–2641
    [Google Scholar]
  7. Broek D., Toda T., Michaeli T., Levin L., Birchmeier C., Zoller M., Powers S., Wigler M. 1987; The Saccharomyces cerevisiae CDC25 gene product regulates the RAS/adenylate cyclase pathway.. Cell 48:789–799
    [Google Scholar]
  8. Camonis J. H., Kalekine M., Gondre B., Garreau H., Boy-Marcotte E., Jacquet M. 1986; Characterization, cloning and sequence analysis of the CDC25 gene which controls the cyclic AMP level of Saccharomyces cerevisiae. EMBO Journal 5:375–380
    [Google Scholar]
  9. Caspani G., Tortora P., Hanozet G. M., Guerritore A. 1985; Glucose-stimulated cAMP increase may be mediated by intracellu-lar acidification in Saccharomyces cerevisiae. FEBS Letters 186:75–79
    [Google Scholar]
  10. Casperson G. F., Walker N., Bourne H. R. 1985; Isolation of the gene encoding adenylate cyclase in Saccharomyces cerevisiae. Proceedings of the National Academy of Sciences of the United States of America 825060–5063
    [Google Scholar]
  11. Celenza J. L., Carlson M. 1984; Cloning and genetic mapping of SNF 1, a gene required for expression of glucose-repressible genes in Saccharomyces cerevisiae. Molecular and Cellular Biology 4:49–53
    [Google Scholar]
  12. Daniel J. 1986; The CDC25 ‘start’ gene of Saccharomyces cerevisiae: sequencing of the active C-terminal fragment and regional homol-ogies with rhodopsin and cytochrome P450.. Current Genetics 10:879–885
    [Google Scholar]
  13. Daniel J., Becker J. M., Enari E., Levitzki A. 1987; The activation of adenylate cyclase by guanyl nucleotides in Saccharomyces cerevisiae is controlled by the CDC25 start gene product.. Molecular and Cellular Biology 7:3857–3861
    [Google Scholar]
  14. Defeo-Jones D., Scolnick E. M., Roller R., Dhar R. 1983; raj-Related gene sequences identified and isolated from Saccharomyces cerevisiae. Nature, London 306:707–709
    [Google Scholar]
  15. Defeo-Jones D., Tatchell K., Robinson L. C., Sigal I. S., Vass W. C., Lowy D. R., Scolnick E. M. 1965; Mammalian and yeast ras gene products: biological function in their heterologous systems.. Science 228:179–184
    [Google Scholar]
  16. Dhar R., Nieto A., Koller R., Defeo-Jones D., Scolnick E. 1984; Nucleotide sequence of two ras H related-genes isolated from the yeast Saccharomyces cerevisiae. Nucleic Acids Research 12:3611–3618
    [Google Scholar]
  17. Entian K.-D. 1986; Glucose repression: a complex regulatory system in yeast. Microbiological Sciences 3:366–371
    [Google Scholar]
  18. Entian K.-D., Zimmermann F. K. 1982; New genes involved in carbon catabolite repression and derepression in the yeast Saccharomyces cerevisiae. Journal of Bacteriology 151:1123–1128
    [Google Scholar]
  19. Gilman A. G. 1987; G proteins: transducers of receptor-generated signals. Annual Review of Biochemistry 56:615–649
    [Google Scholar]
  20. Goldstein A., Lampen J. O. 1975; βd-Fructofuranoside fructohydrolase from yeast. Methods in Enzymology 42C:504–511
    [Google Scholar]
  21. den Hollander J. A., Ugurbil K., Brown T. R., Shulman R. G. 1981; 31P-NMR studies of the effect of oxygen upon glycolysis in yeast. Biochemistry 20:5871–5880
    [Google Scholar]
  22. Ito H., Fukuda Y., Murata K., Kimura A. 1983; Transformation of intact yeast cells treated with alkali cations. Journal of Bacteriology 153:163–168
    [Google Scholar]
  23. Kataoka T., Powers S., Cameron S., Fasano O., Goldfarb M., Broach J., Wigler M. 1985; Functional homology of mammalian and yeast RAS genes.. Cell 40:19–26
    [Google Scholar]
  24. Kataoka T., Powers S., McGill C., Fasano O., Strathern J., Broach J., Wigler M. 1984; Genetic analysis of yeast RAS1 and RAS2 genes.. Cell 37:437–45
    [Google Scholar]
  25. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning; a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  26. Marshall M. S., Gibbs J. B., Scolnick E. M., Sigal I. S. 1987; Regulatory function of the Saccharomyces cerevisiae RAS C-terminus.. Molecular and Cellular Biology 7:2309–2315
    [Google Scholar]
  27. Martegani E., Baroni M., Frascotti G., Alberghina L. 1986a; Molecular cloning and transcriptional analysis of the start gene CDC25 of Saccharomyces cerevisiae. EMBO Journal 5:2363–2369
    [Google Scholar]
  28. Martegani E., Baroni M., Wanoni M. 1986b; Interaction of cAMP with the CDC25-mediated step in the cell cycle of budding yeast. Experimental Cell Research 162:544–548
    [Google Scholar]
  29. Mbonyi K., Thevelein J. M. 1988 The high-affinity glucose
    [Google Scholar]
  30. Mbonyi K., Beullens M., Detremerie K., Geerts L. uptake system is not required for induction of the RAS-mediated cAMP signal by glucose in cells of the yeast Saccharomyces cerevisiae. Biochimica et Biophysica Acta 971:223–226
    [Google Scholar]
  31. 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]
  32. Mbonyi K., Van Aelst L., Arguelles J. C., Jans A. W. H., Thevelein J. M. 1990; Glucose-induced hyperaccumulation of cyclic AMP and defective glucose repression in yeast strains with reduced activity of cyclic AMP-dependent protein kinase. Molecular and Cellular Biology 10:4518–4523
    [Google Scholar]
  33. Munder T., Kuntzel H. 1989; Glucose-induced cAMP signalling in Saccharomyces cerevisiae is mediated by the CDC25 protein.. FEBS Letters 242:341–345
    [Google Scholar]
  34. Nikawa J., Cameron S., Toda T., Ferguson K. M., Wigler M. 1987; Rigorous feedback control of cAMP levels in Saccharomyces cerevisiae. Genes & Development 1:931–937
    [Google Scholar]
  35. Petitjean A., Hilger F., Tatchell K. 1990; Comparison of thermosensitive alleles of the CDC25 gene involved in the cAMP metabolism of Saccharomyces cerevisiae. Genetics 124:797–806
    [Google Scholar]
  36. Portillo F., Mazon M. J. 1986; The Saccharomyces cerevisiae start mutant carrying the cdc25 mutation is defective in activation of plasma membrane ATPase by glucose.. Journal of Bacteriology 168:1254–1257
    [Google Scholar]
  37. Purwin C., Nicolay K., Scheffers W. 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]
  38. Robinson L. C., Gibbs J. B., Marshall M. S., Sigal I. S., Tatchell K. 1987; cdc25: a component of the Ras-adenylate cyclase pathway in Saccharomyces cerevisiae. Science 235:1218–1221
    [Google Scholar]
  39. Sherman F., Fink G. R., Hicks J. B. 1985 Methods in Yeast Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  40. Struhl K., Stinchcomb D. T., Scherer S., Davis R. W. 1979; High-frequency transformation of yeast: autonomous replication of hybrid DNA. Proceedings of the National Academy of Sciences of the United States of America 761035–1039
    [Google Scholar]
  41. Thevelein J. M. 1988; Regulation of trehalase activity by phosphorylation-dephosphorylation during developmental transitions in fungi. Experimental Mycology 12:1–12
    [Google Scholar]
  42. Thevelein J. M., Beullens M. 1985; Cyclic AMP and the stimulation of trehalase activity in the yeast Saccharomyces cerevisiae by carbon sources, nitrogen sources and inhibitors of protein synthesis.. Journal of General Microbiology 131:3199–3209
    [Google Scholar]
  43. Thevelein J. M., Van Laere A. J., Beullens M., Van Assche J. A., Carlier A. R. 1983; Glucose-induced trehalase activation and trehalose mobilization during early germination of Phycomyces blakesleeanus spores.. Journal of General Microbiology 129:719–726
    [Google Scholar]
  44. Thevelein J. M., Beullens M., Honshoven F., Hoebeeck G., Detremerie K., den Hollander J. A., 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]
  45. Thevelein J. M., Beullens M., Honshoven F., Hoebeeck G., Detremerie K., Griewel B., den Hollander J. A., 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]
  46. Toda T., Uno I., Ishikawa T., Powers S., Kataoka T., Broek D., Cameron S., Broach J., Matsumoto K., Wigler M. 1985; In yeast, RAS proteins are controlling elements of adenylate cyclase.. Cell 40:27–36
    [Google Scholar]
  47. Zimmerman F. K., Kaufmann I., Rasenberger H., HauBmann P. 1977; Genetics of carbon catabolite repression in Saccharomyces cerevisiae: genes involved in the derepression process.. Molecular and General Genetics 151:95–103
    [Google Scholar]
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