Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1996 Feb 15;314(Pt 1):95–101. doi: 10.1042/bj3140095

Control of 6-(D-threo-1',2'-dihydroxypropyl) pterin (dictyopterin) synthesis during aggregation of Dictyostelium discoideum. Involvement of the G-protein-linked signalling pathway in the regulation of GTP cyclohydrolase I activity.

M Gütlich 1, K Witter 1, J Bourdais 1, M Veron 1, W Rödl 1, I Ziegler 1
PMCID: PMC1217057  PMID: 8660315

Abstract

6-(D-threo-1',2'-Dihydroxypropylpterin (dictyopterin) has been identified in extracts of growing Dictyostelium dicoideum cells [Klein, Thiery and Tatischeff (1990) Eur. J. Biochem. 187, 665-669]. We demonstrate that it originates from GTP by de novo biosynthesis and that the first committed step is catalysed by GTP cyclohydrolase I, yielding dihydroneopterin triphosphate [neopterin is 6-(D-erythro-1',2',3'-trihydroxypropyl) pterin]. The GTP cyclohydrolase I activity is found in the cytosolic fraction and in a membrane-associated form. The level of a 0.9 kb mRNA coding for GTP cyclohydrolase I decreases to about 10% of its initial value within 2 h after Dictyostelium cells start development induced by starvation. In the cytosolic fraction, the specific activities of GTP cyclohydrolase I, as well as the concentrations of (6R/S)-5,6,7,8-tetrahydrodictyopterin (H4dictyopterin), follow this decline of the mRNA level. In the particulate fraction, however, the specific activities of GTP cyclohydrolase I and, in consequence, H4dictyopterin synthesis, transiently increase and reach a maximum after 4-5 h of development. The time-course of H4dictyopterin concentrations in the starvation medium closely correlates with its production in the membrane fraction. The activity of membrane-associated GTP cyclohydrolase I can be increased by pre-incubation of the cell lysate with guanosine 5'-[gamma-thio]triphosphate and Mg2+. This GTP analogue does not serve as a substrate and has no direct effect on the enzyme activity, indicating that a G-protein-linked signalling pathway is involved in the regulation of GTP cyclohydrolase I activity and thus in H4dictyopterin production during early development of D. discoideum.

Full Text

The Full Text of this article is available as a PDF (476.4 KB).

Selected References

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

  1. Bernstein R. L., Rossier C., van Driel R., Brunner M., Gerisch G. Folate deaminase and cyclic AMP phosphodiesterase in Dictyostelium discoideum: their regulation by extracellular cyclic AMP and folic acid. Cell Differ. 1981 Mar;10(2):79–86. doi: 10.1016/0045-6039(81)90015-4. [DOI] [PubMed] [Google Scholar]
  2. Besmer P. The kit ligand encoded at the murine Steel locus: a pleiotropic growth and differentiation factor. Curr Opin Cell Biol. 1991 Dec;3(6):939–946. doi: 10.1016/0955-0674(91)90111-b. [DOI] [PubMed] [Google Scholar]
  3. Blau N., Niederwieser A. The application of 8-aminoguanosine triphosphate, a new inhibitor of GTP cyclohydrolase I, to the purification of the enzyme from human liver. Biochim Biophys Acta. 1986 Jan 15;880(1):26–31. doi: 10.1016/0304-4165(86)90115-7. [DOI] [PubMed] [Google Scholar]
  4. Bominaar A. A., Molijn A. C., Pestel M., Veron M., Van Haastert P. J. Activation of G-proteins by receptor-stimulated nucleoside diphosphate kinase in Dictyostelium. EMBO J. 1993 Jun;12(6):2275–2279. doi: 10.1002/j.1460-2075.1993.tb05881.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burg A. W., Brown G. M. The biosynthesis of folic acid. 8. Purification and properties of the enzyme that catalyzes the production of formate from carbon atom 8 of guanosine triphosphate. J Biol Chem. 1968 May 10;243(9):2349–2358. [PubMed] [Google Scholar]
  6. Bürki E., Anjard C., Scholder J. C., Reymond C. D. Isolation of two genes encoding putative protein kinases regulated during Dictyostelium discoideum development. Gene. 1991 Jun 15;102(1):57–65. doi: 10.1016/0378-1119(91)90538-m. [DOI] [PubMed] [Google Scholar]
  7. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  8. Devreotes P. N. G protein-linked signaling pathways control the developmental program of Dictyostelium. Neuron. 1994 Feb;12(2):235–241. doi: 10.1016/0896-6273(94)90267-4. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Franke J., Kessin R. A defined minimal medium for axenic strains of Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1977 May;74(5):2157–2161. doi: 10.1073/pnas.74.5.2157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gerisch G. Cyclic AMP and other signals controlling cell development and differentiation in Dictyostelium. Annu Rev Biochem. 1987;56:853–879. doi: 10.1146/annurev.bi.56.070187.004225. [DOI] [PubMed] [Google Scholar]
  12. Gütlich M., Jaeger E., Rücknagel K. P., Werner T., Rödl W., Ziegler I., Bacher A. Human GTP cyclohydrolase I: only one out of three cDNA isoforms gives rise to the active enzyme. Biochem J. 1994 Aug 15;302(Pt 1):215–221. doi: 10.1042/bj3020215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gütlich M., Schott K., Werner T., Bacher A., Ziegler I. Species and tissue specificity of mammalian GTP cyclohydrolase I messenger RNA. Biochim Biophys Acta. 1992 Dec 29;1171(2):133–140. doi: 10.1016/0167-4781(92)90112-d. [DOI] [PubMed] [Google Scholar]
  14. Hadwiger J. A., Lee S., Firtel R. A. The G alpha subunit G alpha 4 couples to pterin receptors and identifies a signaling pathway that is essential for multicellular development in Dictyostelium. Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10566–10570. doi: 10.1073/pnas.91.22.10566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hatakeyama K., Harada T., Suzuki S., Watanabe Y., Kagamiyama H. Purification and characterization of rat liver GTP cyclohydrolase I. Cooperative binding of GTP to the enzyme. J Biol Chem. 1989 Dec 25;264(36):21660–21664. [PubMed] [Google Scholar]
  16. Heine M. C., Brown G. M. Enzymatic epimerization of D-erythro-dihydroneopterin triphosphate to L-threo-dihydroneopterin triphosphate. Biochim Biophys Acta. 1975 Dec 5;411(2):236–249. doi: 10.1016/0304-4165(75)90304-9. [DOI] [PubMed] [Google Scholar]
  17. Hori H., Osawa S., Iwabuchi M. The nucleotide sequence of 5S rRNA from a cellular slime mold Dictyostelium discoideum. Nucleic Acids Res. 1980 Dec 11;8(23):5535–5539. doi: 10.1093/nar/8.23.5535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Huber C., Batchelor J. R., Fuchs D., Hausen A., Lang A., Niederwieser D., Reibnegger G., Swetly P., Troppmair J., Wachter H. Immune response-associated production of neopterin. Release from macrophages primarily under control of interferon-gamma. J Exp Med. 1984 Jul 1;160(1):310–316. doi: 10.1084/jem.160.1.310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kerler F., Hültner L., Ziegler I., Katzenmaier G., Bacher A. Analysis of the tetrahydrobiopterin synthesizing system during maturation of murine reticulocytes. J Cell Physiol. 1990 Feb;142(2):268–271. doi: 10.1002/jcp.1041420208. [DOI] [PubMed] [Google Scholar]
  20. Kerler F., Schwarzkopf B., Katzenmaier G., Le Van Q., Schmid C., Ziegler I., Bacher A. Biosynthesis of tetrahydrobiopterin: a sensitive assay of 6-pyruvoyltetrahydropterin synthase using [2'-3H]dihydroneopterin 3'-triphosphate as substrate. Biochim Biophys Acta. 1989 Jan 27;990(1):15–17. doi: 10.1016/s0304-4165(89)80005-4. [DOI] [PubMed] [Google Scholar]
  21. Klein R., Tatischeff I., Tham G., Mano N. Chiral lumazines: preparation, properties, enantiomeric separation. Chirality. 1994;6(7):564–571. doi: 10.1002/chir.530060709. [DOI] [PubMed] [Google Scholar]
  22. Klein R., Thiery R., Tatischeff I. Dictyopterin, 6-(D-threo-1,2-dihydroxypropyl)-pterin, a new natural isomer of L-biopterin. Isolation from vegetative cells of Dictyostelium discoideum and identification. Eur J Biochem. 1990 Feb 14;187(3):665–669. doi: 10.1111/j.1432-1033.1990.tb15351.x. [DOI] [PubMed] [Google Scholar]
  23. Maier J., Witter K., Gütlich M., Ziegler I., Werner T., Ninnemann H. Homology cloning of GTP-cyclohydrolase I from various unrelated eukaryotes by reverse-transcription polymerase chain reaction using a general set of degenerate primers. Biochem Biophys Res Commun. 1995 Jul 17;212(2):705–711. doi: 10.1006/bbrc.1995.2026. [DOI] [PubMed] [Google Scholar]
  24. Maier J., Ziegler I. Purification and properties of human sepiapterin reductase from placenta. Adv Exp Med Biol. 1993;338:199–202. doi: 10.1007/978-1-4615-2960-6_40. [DOI] [PubMed] [Google Scholar]
  25. Nichol C. A., Smith G. K., Duch D. S. Biosynthesis and metabolism of tetrahydrobiopterin and molybdopterin. Annu Rev Biochem. 1985;54:729–764. doi: 10.1146/annurev.bi.54.070185.003501. [DOI] [PubMed] [Google Scholar]
  26. Pan P., Hall E. M., Bonner J. T. Determination of the active portion of the folic acid molecule in cellular slime mold chemotaxis. J Bacteriol. 1975 Apr;122(1):185–191. doi: 10.1128/jb.122.1.185-191.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Pan P., Hall E. M., Bonner J. T. Folic acid as second chemotactic substance in the cellular slime moulds. Nat New Biol. 1972 Jun 7;237(75):181–182. doi: 10.1038/newbio237181a0. [DOI] [PubMed] [Google Scholar]
  28. Pan P., Wurster B. Inactivation of the chemoattractant folic acid by cellular slime molds and identification of the reaction product. J Bacteriol. 1978 Dec;136(3):955–959. doi: 10.1128/jb.136.3.955-959.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schott K., Gütlich M., Ziegler I. Induction of GTP-cyclohydrolase I mRNA expression by lectin activation and interferon-gamma treatment in human cells associated with the immune response. J Cell Physiol. 1993 Jul;156(1):12–16. doi: 10.1002/jcp.1041560103. [DOI] [PubMed] [Google Scholar]
  30. Snaar-Jagalska B. E., Van Haastert P. J. G-protein assays in Dictyostelium. Methods Enzymol. 1994;237:387–408. doi: 10.1016/s0076-6879(94)37077-x. [DOI] [PubMed] [Google Scholar]
  31. Sussman M. Cultivation and synchronous morphogenesis of Dictyostelium under controlled experimental conditions. Methods Cell Biol. 1987;28:9–29. doi: 10.1016/s0091-679x(08)61635-0. [DOI] [PubMed] [Google Scholar]
  32. Tillinghast H. S., Newell P. C. Chemotaxis towards pteridines during development of Dictyostelium. J Cell Sci. 1987 Feb;87(Pt 1):45–53. doi: 10.1242/jcs.87.1.45. [DOI] [PubMed] [Google Scholar]
  33. Van Haastert P. J., De Wit R. J., Grijpma Y., Konijn T. M. Identification of a pterin as the acrasin of the cellular slime mold Dictyostelium lacteum. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6270–6274. doi: 10.1073/pnas.79.20.6270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Van Haastert P. J., Janssens P. M., Erneux C. Sensory transduction in eukaryotes. A comparison between Dictyostelium and vertebrate cells. Eur J Biochem. 1991 Jan 30;195(2):289–303. doi: 10.1111/j.1432-1033.1991.tb15706.x. [DOI] [PubMed] [Google Scholar]
  35. Wachter H., Hausen A., Reider E., Schweiger M. Pteridine excretion from cells as indicator of cell proliferation. Naturwissenschaften. 1980 Dec;67(12):610–611. doi: 10.1007/BF00396550. [DOI] [PubMed] [Google Scholar]
  36. Watts D. J., Ashworth J. M. Growth of myxameobae of the cellular slime mould Dictyostelium discoideum in axenic culture. Biochem J. 1970 Sep;119(2):171–174. doi: 10.1042/bj1190171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Werner-Felmayer G., Golderer G., Werner E. R., Gröbner P., Wachter H. Pteridine biosynthesis and nitric oxide synthase in Physarum polycephalum. Biochem J. 1994 Nov 15;304(Pt 1):105–111. doi: 10.1042/bj3040105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wurster B., Bek F., Butz U. Folic acid and pterin deaminases in Dictyostelium discoideum: kinetic properties and regulation by folic acid, pterin, and adenosine 3',5'-phosphate. J Bacteriol. 1981 Oct;148(1):183–192. doi: 10.1128/jb.148.1.183-192.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Ziegler I., Hültner L., Egger D., Kempkes B., Mailhammer R., Gillis S., Rödl W. In a concerted action kit ligand and interleukin 3 control the synthesis of serotonin in murine bone marrow-derived mast cells. Up-regulation of GTP cyclohydrolase I and tryptophan 5-monooxygenase activity by the kit ligand. J Biol Chem. 1993 Jun 15;268(17):12544–12551. [PubMed] [Google Scholar]
  40. Ziegler I. Pteridine formation during lectin-induced lymphocyte activation. J Cell Biochem. 1985;28(3):197–206. doi: 10.1002/jcb.240280303. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES