Skip to main content
NCBI home page
Genetics logoLink to Genetics
. 2000 Apr;154(4):1463–1471. doi: 10.1093/genetics/154.4.1463

The fission yeast git5 gene encodes a Gbeta subunit required for glucose-triggered adenylate cyclase activation.

S Landry 1, M T Pettit 1, E Apolinario 1, C S Hoffman 1
PMCID: PMC1461029  PMID: 10747045

Abstract

Fission yeast adenylate cyclase is activated by the gpa2 Galpha subunit of a heterotrimeric guanine-nucleotide binding protein (G protein). We show that the git5 gene, also required for this activation, encodes a Gbeta subunit. In contrast to another study, we show that git5 is not a negative regulator of the gpa1 Galpha involved in the pheromone response pathway. While 43% identical to mammalian Gbeta's, the git5 protein lacks the amino-terminal coiled-coil found in other Gbeta subunits, yet the gene possesses some of the coding capacity for this structure 5' to its ORF. Although both gpa2 (Galpha) and git5 (Gbeta) are required for adenylate cyclase activation, only gpa2 is needed to maintain basal cAMP levels. Strains bearing a git5 disruption are derepressed for fbp1 transcription and sexual development even while growing in a glucose-rich environment, although fbp1 derepression is half that observed in gpa2 deletion strains. Multicopy gpa2 partially suppresses the loss of git5, while the converse is not true. These data suggest that Gbeta is required for activation of adenylate cyclase either by promoting the activation of Galpha or by independently activating adenylate cyclase subsequent to Galpha stimulation as seen in type II mammalian adenylate cyclase activation.

Full Text

The Full Text of this article is available as a PDF (344.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Apolinario E., Nocero M., Jin M., Hoffman C. S. Cloning and manipulation of the Schizosaccharomyces pombe his7+ gene as a new selectable marker for molecular genetic studies. Curr Genet. 1993 Dec;24(6):491–495. doi: 10.1007/BF00351711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Clapham D. E., Neer E. J. New roles for G-protein beta gamma-dimers in transmembrane signalling. Nature. 1993 Sep 30;365(6445):403–406. doi: 10.1038/365403a0. [DOI] [PubMed] [Google Scholar]
  3. Dal Santo P., Blanchard B., Hoffman C. S. The Schizosaccharomyces pombe pyp1 protein tyrosine phosphatase negatively regulates nutrient monitoring pathways. J Cell Sci. 1996 Jul;109(Pt 7):1919–1925. doi: 10.1242/jcs.109.7.1919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Federman A. D., Conklin B. R., Schrader K. A., Reed R. R., Bourne H. R. Hormonal stimulation of adenylyl cyclase through Gi-protein beta gamma subunits. Nature. 1992 Mar 12;356(6365):159–161. doi: 10.1038/356159a0. [DOI] [PubMed] [Google Scholar]
  5. Gao B. N., Gilman A. G. Cloning and expression of a widely distributed (type IV) adenylyl cyclase. Proc Natl Acad Sci U S A. 1991 Nov 15;88(22):10178–10182. doi: 10.1073/pnas.88.22.10178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Garcia-Higuera I., Fenoglio J., Li Y., Lewis C., Panchenko M. P., Reiner O., Smith T. F., Neer E. J. Folding of proteins with WD-repeats: comparison of six members of the WD-repeat superfamily to the G protein beta subunit. Biochemistry. 1996 Nov 5;35(44):13985–13994. doi: 10.1021/bi9612879. [DOI] [PubMed] [Google Scholar]
  7. Garritsen A., van Galen P. J., Simonds W. F. The N-terminal coiled-coil domain of beta is essential for gamma association: a model for G-protein beta gamma subunit interaction. Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7706–7710. doi: 10.1073/pnas.90.16.7706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gilman A. G. G proteins and dual control of adenylate cyclase. Cell. 1984 Mar;36(3):577–579. doi: 10.1016/0092-8674(84)90336-2. [DOI] [PubMed] [Google Scholar]
  9. Gilman A. G. G proteins: transducers of receptor-generated signals. Annu Rev Biochem. 1987;56:615–649. doi: 10.1146/annurev.bi.56.070187.003151. [DOI] [PubMed] [Google Scholar]
  10. Hoffman C. S., Winston F. Glucose repression of transcription of the Schizosaccharomyces pombe fbp1 gene occurs by a cAMP signaling pathway. Genes Dev. 1991 Apr;5(4):561–571. doi: 10.1101/gad.5.4.561. [DOI] [PubMed] [Google Scholar]
  11. Hoffman C. S., Winston F. Isolation and characterization of mutants constitutive for expression of the fbp1 gene of Schizosaccharomyces pombe. Genetics. 1990 Apr;124(4):807–816. doi: 10.1093/genetics/124.4.807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Isshiki T., Mochizuki N., Maeda T., Yamamoto M. Characterization of a fission yeast gene, gpa2, that encodes a G alpha subunit involved in the monitoring of nutrition. Genes Dev. 1992 Dec;6(12B):2455–2462. doi: 10.1101/gad.6.12b.2455. [DOI] [PubMed] [Google Scholar]
  13. Jin M., Fujita M., Culley B. M., Apolinario E., Yamamoto M., Maundrell K., Hoffman C. S. sck1, a high copy number suppressor of defects in the cAMP-dependent protein kinase pathway in fission yeast, encodes a protein homologous to the Saccharomyces cerevisiae SCH9 kinase. Genetics. 1995 Jun;140(2):457–467. doi: 10.1093/genetics/140.2.457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kawamukai M., Ferguson K., Wigler M., Young D. Genetic and biochemical analysis of the adenylyl cyclase of Schizosaccharomyces pombe. Cell Regul. 1991 Feb;2(2):155–164. doi: 10.1091/mbc.2.2.155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kim D. U., Park S. K., Chung K. S., Choi M. U., Yoo H. S. The G protein beta subunit Gpb1 of Schizosaccharomyces pombe is a negative regulator of sexual development. Mol Gen Genet. 1996 Aug 27;252(1-2):20–32. doi: 10.1007/BF02173201. [DOI] [PubMed] [Google Scholar]
  16. Kohli J. Genetic nomenclature and gene list of the fission yeast Schizosaccharomyces pombe. Curr Genet. 1987;11(8):575–589. doi: 10.1007/BF00393919. [DOI] [PubMed] [Google Scholar]
  17. Lambright D. G., Sondek J., Bohm A., Skiba N. P., Hamm H. E., Sigler P. B. The 2.0 A crystal structure of a heterotrimeric G protein. Nature. 1996 Jan 25;379(6563):311–319. doi: 10.1038/379311a0. [DOI] [PubMed] [Google Scholar]
  18. Levitzki A., Bar-Sinai A. The regulation of adenylyl cyclase by receptor-operated G proteins. Pharmacol Ther. 1991;50(3):271–283. doi: 10.1016/0163-7258(91)90045-n. [DOI] [PubMed] [Google Scholar]
  19. Maeda T., Mochizuki N., Yamamoto M. Adenylyl cyclase is dispensable for vegetative cell growth in the fission yeast Schizosaccharomyces pombe. Proc Natl Acad Sci U S A. 1990 Oct;87(20):7814–7818. doi: 10.1073/pnas.87.20.7814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Maeda T., Watanabe Y., Kunitomo H., Yamamoto M. Cloning of the pka1 gene encoding the catalytic subunit of the cAMP-dependent protein kinase in Schizosaccharomyces pombe. J Biol Chem. 1994 Apr 1;269(13):9632–9637. [PubMed] [Google Scholar]
  21. Mochizuki N., Yamamoto M. Reduction in the intracellular cAMP level triggers initiation of sexual development in fission yeast. Mol Gen Genet. 1992 May;233(1-2):17–24. doi: 10.1007/BF00587556. [DOI] [PubMed] [Google Scholar]
  22. Molz L., Booher R., Young P., Beach D. cdc2 and the regulation of mitosis: six interacting mcs genes. Genetics. 1989 Aug;122(4):773–782. doi: 10.1093/genetics/122.4.773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nocero M., Isshiki T., Yamamoto M., Hoffman C. S. Glucose repression of fbp1 transcription of Schizosaccharomyces pombe is partially regulated by adenylate cyclase activation by a G protein alpha subunit encoded by gpa2 (git8). Genetics. 1994 Sep;138(1):39–45. doi: 10.1093/genetics/138.1.39. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nomoto S., Nakayama N., Arai K., Matsumoto K. Regulation of the yeast pheromone response pathway by G protein subunits. EMBO J. 1990 Mar;9(3):691–696. doi: 10.1002/j.1460-2075.1990.tb08161.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Obara T., Nakafuku M., Yamamoto M., Kaziro Y. Isolation and characterization of a gene encoding a G-protein alpha subunit from Schizosaccharomyces pombe: involvement in mating and sporulation pathways. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5877–5881. doi: 10.1073/pnas.88.13.5877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pellegrino S., Zhang S., Garritsen A., Simonds W. F. The coiled-coil region of the G protein beta subunit. Mutational analysis of Ggamma and effector interactions. J Biol Chem. 1997 Oct 3;272(40):25360–25366. doi: 10.1074/jbc.272.40.25360. [DOI] [PubMed] [Google Scholar]
  27. Pidoux A. L., LeDizet M., Cande W. Z. Fission yeast pkl1 is a kinesin-related protein involved in mitotic spindle function. Mol Biol Cell. 1996 Oct;7(10):1639–1655. doi: 10.1091/mbc.7.10.1639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Simon M. I., Strathmann M. P., Gautam N. Diversity of G proteins in signal transduction. Science. 1991 May 10;252(5007):802–808. doi: 10.1126/science.1902986. [DOI] [PubMed] [Google Scholar]
  29. Sternweis P. C. The active role of beta gamma in signal transduction. Curr Opin Cell Biol. 1994 Apr;6(2):198–203. doi: 10.1016/0955-0674(94)90136-8. [DOI] [PubMed] [Google Scholar]
  30. Tang W. J., Gilman A. G. Type-specific regulation of adenylyl cyclase by G protein beta gamma subunits. Science. 1991 Dec 6;254(5037):1500–1503. doi: 10.1126/science.1962211. [DOI] [PubMed] [Google Scholar]
  31. Thompson J. D., Higgins D. G., Gibson T. J. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994 Nov 11;22(22):4673–4680. doi: 10.1093/nar/22.22.4673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wall M. A., Coleman D. E., Lee E., Iñiguez-Lluhi J. A., Posner B. A., Gilman A. G., Sprang S. R. The structure of the G protein heterotrimer Gi alpha 1 beta 1 gamma 2. Cell. 1995 Dec 15;83(6):1047–1058. doi: 10.1016/0092-8674(95)90220-1. [DOI] [PubMed] [Google Scholar]
  33. Watanabe Y., Lino Y., Furuhata K., Shimoda C., Yamamoto M. The S.pombe mei2 gene encoding a crucial molecule for commitment to meiosis is under the regulation of cAMP. EMBO J. 1988 Mar;7(3):761–767. doi: 10.1002/j.1460-2075.1988.tb02873.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Whiteway M., Hougan L., Dignard D., Bell L., Saari G., Grant F., O'Hara P., MacKay V. L., Thomas D. Y. Function of the STE4 and STE18 genes in mating pheromone signal transduction in Saccharomyces cerevisiae. Cold Spring Harb Symp Quant Biol. 1988;53(Pt 2):585–590. doi: 10.1101/sqb.1988.053.01.067. [DOI] [PubMed] [Google Scholar]
  35. Yamawaki-Kataoka Y., Tamaoki T., Choe H. R., Tanaka H., Kataoka T. Adenylate cyclases in yeast: a comparison of the genes from Schizosaccharomyces pombe and Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5693–5697. doi: 10.1073/pnas.86.15.5693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Young D., Riggs M., Field J., Vojtek A., Broek D., Wigler M. The adenylyl cyclase gene from Schizosaccharomyces pombe. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7989–7993. doi: 10.1073/pnas.86.20.7989. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES