Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1991 Sep 15;88(18):8213–8217. doi: 10.1073/pnas.88.18.8213

Identification of Dictyostelium G alpha genes expressed during multicellular development.

J A Hadwiger 1, T M Wilkie 1, M Strathmann 1, R A Firtel 1
PMCID: PMC52477  PMID: 1910174

Abstract

Guanine nucleotide-binding protein (G protein)-mediated signal transduction constitutes a common mechanism by which cells receive and respond to a diverse set of environmental signals. Many of the signals involved in the developmental life cycle of the slime mold Dictyostelium have been postulated to be transduced by such pathways and, in some cases, these pathways have been demonstrated to be dependent on specific G proteins. Using the polymerase chain reaction, we have identified two additional Dictyostelium G alpha genes, G alpha 4 and G alpha 5, that are developmentally regulated. Transcripts from both of these genes are primarily expressed during the multicellular stages of development, suggesting possible roles in cell differentiation or morphogenesis. The entire G alpha 4 gene was sequenced and found to encode a protein consisting of 345 amino acids. The G alpha 4 subunit is homologous to other previously identified G alpha subunits, including the Dictyostelium G alpha 1 (43% identity) and G alpha 2 (41% identity) subunits. However, the G alpha 4 subunit contains some unusual sequence divergences in residues highly conserved among most eukaryotic G alpha subunits, suggesting that G alpha 4 may be a member of another class of G alpha subunits.

Full text

PDF
8217

Images in this article

8215Image
on p.8215

Selected References

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

  1. Amatruda T. T., 3rd, Steele D. A., Slepak V. Z., Simon M. I. G alpha 16, a G protein alpha subunit specifically expressed in hematopoietic cells. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5587–5591. doi: 10.1073/pnas.88.13.5587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bloom L., Kay R. R. The search for morphogens in Dictyostelium. Bioessays. 1988 Dec;9(6):187–191. doi: 10.1002/bies.950090602. [DOI] [PubMed] [Google Scholar]
  3. Casey P. J., Fong H. K., Simon M. I., Gilman A. G. Gz, a guanine nucleotide-binding protein with unique biochemical properties. J Biol Chem. 1990 Feb 5;265(4):2383–2390. [PubMed] [Google Scholar]
  4. Chou P. Y., Fasman G. D. Prediction of protein conformation. Biochemistry. 1974 Jan 15;13(2):222–245. doi: 10.1021/bi00699a002. [DOI] [PubMed] [Google Scholar]
  5. Clack J. W., Oakley B., 2nd, Stein P. J. Injection of GTP-binding protein or cyclic GMP phosphodiesterase hyperpolarizes retinal rods. Nature. 1983 Sep 1;305(5929):50–52. doi: 10.1038/305050a0. [DOI] [PubMed] [Google Scholar]
  6. De Wit R. J., Bulgakov R. Guanine nucleotides modulate the ligand binding properties of cell surface folate receptors in Dictyostelium discoideum. FEBS Lett. 1985 Jan 7;179(2):257–261. doi: 10.1016/0014-5793(85)80530-5. [DOI] [PubMed] [Google Scholar]
  7. Der C. J., Finkel T., Cooper G. M. Biological and biochemical properties of human rasH genes mutated at codon 61. Cell. 1986 Jan 17;44(1):167–176. doi: 10.1016/0092-8674(86)90495-2. [DOI] [PubMed] [Google Scholar]
  8. Devreotes P. Dictyostelium discoideum: a model system for cell-cell interactions in development. Science. 1989 Sep 8;245(4922):1054–1058. doi: 10.1126/science.2672337. [DOI] [PubMed] [Google Scholar]
  9. Dynes J. L., Firtel R. A. Molecular complementation of a genetic marker in Dictyostelium using a genomic DNA library. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7966–7970. doi: 10.1073/pnas.86.20.7966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Esch R. K., Firtel R. A. cAMP and cell sorting control the spatial expression of a developmentally essential cell-type-specific ras gene in Dictyostelium. Genes Dev. 1991 Jan;5(1):9–21. doi: 10.1101/gad.5.1.9. [DOI] [PubMed] [Google Scholar]
  11. Firtel R. A., van Haastert P. J., Kimmel A. R., Devreotes P. N. G protein linked signal transduction pathways in development: dictyostelium as an experimental system. Cell. 1989 Jul 28;58(2):235–239. doi: 10.1016/0092-8674(89)90837-4. [DOI] [PubMed] [Google Scholar]
  12. Fong H. K., Yoshimoto K. K., Eversole-Cire P., Simon M. I. Identification of a GTP-binding protein alpha subunit that lacks an apparent ADP-ribosylation site for pertussis toxin. Proc Natl Acad Sci U S A. 1988 May;85(9):3066–3070. doi: 10.1073/pnas.85.9.3066. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Ginsburg G., Kimmel A. R. Inositol trisphosphate and diacylglycerol can differentially modulate gene expression in Dictyostelium. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9332–9336. doi: 10.1073/pnas.86.23.9332. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gomer R. H., Armstrong D., Leichtling B. H., Firtel R. A. cAMP induction of prespore and prestalk gene expression in Dictyostelium is mediated by the cell-surface cAMP receptor. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8624–8628. doi: 10.1073/pnas.83.22.8624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Haberstroh L., Firtel R. A. A spatial gradient of expression of a cAMP-regulated prespore cell-type-specific gene in Dictyostelium. Genes Dev. 1990 Apr;4(4):596–612. doi: 10.1101/gad.4.4.596. [DOI] [PubMed] [Google Scholar]
  18. Halliday K. R. Regional homology in GTP-binding proto-oncogene products and elongation factors. J Cyclic Nucleotide Protein Phosphor Res. 1983;9(6):435–448. [PubMed] [Google Scholar]
  19. Haribabu B., Dottin R. P. Pharmacological characterization of cyclic AMP receptors mediating gene regulation in Dictyostelium discoideum. Mol Cell Biol. 1986 Jul;6(7):2402–2408. doi: 10.1128/mcb.6.7.2402. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Holbrook S. R., Kim S. H. Molecular model of the G protein alpha subunit based on the crystal structure of the HRAS protein. Proc Natl Acad Sci U S A. 1989 Mar;86(6):1751–1755. doi: 10.1073/pnas.86.6.1751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hopp T. P., Woods K. R. Prediction of protein antigenic determinants from amino acid sequences. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3824–3828. doi: 10.1073/pnas.78.6.3824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Janssens P. M., Van Haastert P. J. Molecular basis of transmembrane signal transduction in Dictyostelium discoideum. Microbiol Rev. 1987 Dec;51(4):396–418. doi: 10.1128/mr.51.4.396-418.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Johnson G. L., Dhanasekaran N. The G-protein family and their interaction with receptors. Endocr Rev. 1989 Aug;10(3):317–331. doi: 10.1210/edrv-10-3-317. [DOI] [PubMed] [Google Scholar]
  24. Kesbeke F., Snaar-Jagalska B. E., Van Haastert P. J. Signal transduction in Dictyostelium fgd A mutants with a defective interaction between surface cAMP receptors and a GTP-binding regulatory protein. J Cell Biol. 1988 Aug;107(2):521–528. doi: 10.1083/jcb.107.2.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kimmel A. R. Different molecular mechanisms for cAMP regulation of gene expression during Dictyostelium development. Dev Biol. 1987 Jul;122(1):163–171. doi: 10.1016/0012-1606(87)90342-3. [DOI] [PubMed] [Google Scholar]
  26. Kimmel A. R., Firtel R. A. Sequence organization in Dictyostelium: unique structure at the 5'-ends of protein coding genes. Nucleic Acids Res. 1983 Jan 25;11(2):541–552. doi: 10.1093/nar/11.2.541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Klein P. S., Sun T. J., Saxe C. L., 3rd, Kimmel A. R., Johnson R. L., Devreotes P. N. A chemoattractant receptor controls development in Dictyostelium discoideum. Science. 1988 Sep 16;241(4872):1467–1472. doi: 10.1126/science.3047871. [DOI] [PubMed] [Google Scholar]
  28. Kumagai A., Hadwiger J. A., Pupillo M., Firtel R. A. Molecular genetic analysis of two G alpha protein subunits in Dictyostelium. J Biol Chem. 1991 Jan 15;266(2):1220–1228. [PubMed] [Google Scholar]
  29. Kumagai A., Pupillo M., Gundersen R., Miake-Lye R., Devreotes P. N., Firtel R. A. Regulation and function of G alpha protein subunits in Dictyostelium. Cell. 1989 Apr 21;57(2):265–275. doi: 10.1016/0092-8674(89)90964-1. [DOI] [PubMed] [Google Scholar]
  30. Landis C. A., Masters S. B., Spada A., Pace A. M., Bourne H. R., Vallar L. GTPase inhibiting mutations activate the alpha chain of Gs and stimulate adenylyl cyclase in human pituitary tumours. Nature. 1989 Aug 31;340(6236):692–696. doi: 10.1038/340692a0. [DOI] [PubMed] [Google Scholar]
  31. Lochrie M. A., Simon M. I. G protein multiplicity in eukaryotic signal transduction systems. Biochemistry. 1988 Jul 12;27(14):4957–4965. doi: 10.1021/bi00414a001. [DOI] [PubMed] [Google Scholar]
  32. Lyons J., Landis C. A., Harsh G., Vallar L., Grünewald K., Feichtinger H., Duh Q. Y., Clark O. H., Kawasaki E., Bourne H. R. Two G protein oncogenes in human endocrine tumors. Science. 1990 Aug 10;249(4969):655–659. doi: 10.1126/science.2116665. [DOI] [PubMed] [Google Scholar]
  33. Ma H., Yanofsky M. F., Meyerowitz E. M. Molecular cloning and characterization of GPA1, a G protein alpha subunit gene from Arabidopsis thaliana. Proc Natl Acad Sci U S A. 1990 May;87(10):3821–3825. doi: 10.1073/pnas.87.10.3821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Mann S. K., Firtel R. A. Cyclic AMP regulation of early gene expression in Dictyostelium discoideum: mediation via the cell surface cyclic AMP receptor. Mol Cell Biol. 1987 Jan;7(1):458–469. doi: 10.1128/mcb.7.1.458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Masters S. B., Stroud R. M., Bourne H. R. Family of G protein alpha chains: amphipathic analysis and predicted structure of functional domains. Protein Eng. 1986 Oct-Nov;1(1):47–54. [PubMed] [Google Scholar]
  36. Matsuoka M., Itoh H., Kozasa T., Kaziro Y. Sequence analysis of cDNA and genomic DNA for a putative pertussis toxin-insensitive guanine nucleotide-binding regulatory protein alpha subunit. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5384–5388. doi: 10.1073/pnas.85.15.5384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. May D. C., Ross E. M., Gilman A. G., Smigel M. D. Reconstitution of catecholamine-stimulated adenylate cyclase activity using three purified proteins. J Biol Chem. 1985 Dec 15;260(29):15829–15833. [PubMed] [Google Scholar]
  38. Mehdy M. C., Firtel R. A. A secreted factor and cyclic AMP jointly regulate cell-type-specific gene expression in Dictyostelium discoideum. Mol Cell Biol. 1985 Apr;5(4):705–713. doi: 10.1128/mcb.5.4.705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Miyajima I., Arai K., Matsumoto K. GPA1Val-50 mutation in the mating-factor signaling pathway in Saccharomyces cerevisiae. Mol Cell Biol. 1989 Jun;9(6):2289–2297. doi: 10.1128/mcb.9.6.2289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Nakafuku M., Obara T., Kaibuchi K., Miyajima I., Miyajima A., Itoh H., Nakamura S., Arai K., Matsumoto K., Kaziro Y. Isolation of a second yeast Saccharomyces cerevisiae gene (GPA2) coding for guanine nucleotide-binding regulatory protein: studies on its structure and possible functions. Proc Natl Acad Sci U S A. 1988 Mar;85(5):1374–1378. doi: 10.1073/pnas.85.5.1374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Oyama M., Blumberg D. D. Interaction of cAMP with the cell-surface receptor induces cell-type-specific mRNA accumulation in Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4819–4823. doi: 10.1073/pnas.83.13.4819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Pang I. H., Sternweis P. C. Isolation of the alpha subunits of GTP-binding regulatory proteins by affinity chromatography with immobilized beta gamma subunits. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7814–7818. doi: 10.1073/pnas.86.20.7814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Pang I. H., Sternweis P. C. Purification of unique alpha subunits of GTP-binding regulatory proteins (G proteins) by affinity chromatography with immobilized beta gamma subunits. J Biol Chem. 1990 Oct 25;265(30):18707–18712. [PubMed] [Google Scholar]
  44. Pupillo M., Kumagai A., Pitt G. S., Firtel R. A., Devreotes P. N. Multiple alpha subunits of guanine nucleotide-binding proteins in Dictyostelium. Proc Natl Acad Sci U S A. 1989 Jul;86(13):4892–4896. doi: 10.1073/pnas.86.13.4892. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Ross E. M. Signal sorting and amplification through G protein-coupled receptors. Neuron. 1989 Aug;3(2):141–152. doi: 10.1016/0896-6273(89)90027-5. [DOI] [PubMed] [Google Scholar]
  46. Saxe C. L., 3rd, Johnson R. L., Devreotes P. N., Kimmel A. R. Expression of a cAMP receptor gene of Dictyostelium and evidence for a multigene family. Genes Dev. 1991 Jan;5(1):1–8. doi: 10.1101/gad.5.1.1. [DOI] [PubMed] [Google Scholar]
  47. Saxe C. L., 3rd, Johnson R., Devreotes P. N., Kimmel A. R. Multiple genes for cell surface cAMP receptors in Dictyostelium discoideum. Dev Genet. 1991;12(1-2):6–13. doi: 10.1002/dvg.1020120104. [DOI] [PubMed] [Google Scholar]
  48. Schaap P., Wang M. Interactions between adenosine and oscillatory cAMP signaling regulate size and pattern in Dictyostelium. Cell. 1986 Apr 11;45(1):137–144. doi: 10.1016/0092-8674(86)90545-3. [DOI] [PubMed] [Google Scholar]
  49. Schaap P., van Driel R. V. Induction of post-aggregative differentiation in Dictyostelium discoideum by cAMP. Evidence of involvement of the cell surface cAMP receptor. Exp Cell Res. 1985 Aug;159(2):388–398. doi: 10.1016/s0014-4827(85)80012-4. [DOI] [PubMed] [Google Scholar]
  50. Strathmann M., Simon M. I. G protein diversity: a distinct class of alpha subunits is present in vertebrates and invertebrates. Proc Natl Acad Sci U S A. 1990 Dec;87(23):9113–9117. doi: 10.1073/pnas.87.23.9113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Strathmann M., Wilkie T. M., Simon M. I. Diversity of the G-protein family: sequences from five additional alpha subunits in the mouse. Proc Natl Acad Sci U S A. 1989 Oct;86(19):7407–7409. doi: 10.1073/pnas.86.19.7407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Stryer L., Bourne H. R. G proteins: a family of signal transducers. Annu Rev Cell Biol. 1986;2:391–419. doi: 10.1146/annurev.cb.02.110186.002135. [DOI] [PubMed] [Google Scholar]
  53. Sun T. J., Van Haastert P. J., Devreotes P. N. Surface cAMP receptors mediate multiple responses during development in Dictyostelium: evidenced by antisense mutagenesis. J Cell Biol. 1990 May;110(5):1549–1554. doi: 10.1083/jcb.110.5.1549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Theibert A., Devreotes P. N. Surface receptor-mediated activation of adenylate cyclase in Dictyostelium. Regulation by guanine nucleotides in wild-type cells and aggregation deficient mutants. J Biol Chem. 1986 Nov 15;261(32):15121–15125. [PubMed] [Google Scholar]
  55. Van Haastert P. J., De Vries M. J., Penning L. C., Roovers E., Van der Kaay J., Erneux C., Van Lookeren Campagne M. M. Chemoattractant and guanosine 5'-[gamma-thio]triphosphate induce the accumulation of inositol 1,4,5-trisphosphate in Dictyostelium cells that are labelled with [3H]inositol by electroporation. Biochem J. 1989 Mar 1;258(2):577–586. doi: 10.1042/bj2580577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Van Haastert P. J., Snaar-Jagalska B. E., Janssens P. M. The regulation of adenylate cyclase by guanine nucleotides in Dictyostelium discoideum membranes. Eur J Biochem. 1987 Jan 15;162(2):251–258. doi: 10.1111/j.1432-1033.1987.tb10592.x. [DOI] [PubMed] [Google Scholar]
  57. West R. E., Jr, Moss J., Vaughan M., Liu T., Liu T. Y. Pertussis toxin-catalyzed ADP-ribosylation of transducin. Cysteine 347 is the ADP-ribose acceptor site. J Biol Chem. 1985 Nov 25;260(27):14428–14430. [PubMed] [Google Scholar]
  58. Woon C. W., Heasley L., Osawa S., Johnson G. L. Mutation of glycine 49 to valine in the alpha subunit of GS results in the constitutive elevation of cyclic AMP synthesis. Biochemistry. 1989 May 30;28(11):4547–4551. doi: 10.1021/bi00437a006. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES