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. 1994 Jan 1;297(Pt 1):181–187. doi: 10.1042/bj2970181

Phospholipase C in Dictyostelium discoideum. Cyclic AMP surface receptor and G-protein-regulated activity in vitro.

A A Bominaar 1, F Kesbeke 1, P J Van Haastert 1
PMCID: PMC1137808  PMID: 8280097

Abstract

The cellular slime mould Dictyostelium discoideum shows several responses after stimulation with the chemoattractant cAMP, including a transient rise in cyclic AMP (cAMP), cGMP and Ins(1,4,5)P3. In this paper the regulation of phospholipase C in vitro is described. Under our experimental conditions commercial PtdIns(4,5)P2 cannot be used to analyse phospholipase C activity in Dictyostelium lysates, because it is hydrolysed mainly to glycerophosphoinositol instead of Ins(1,4,5)P3. Enzyme activity was determined with endogenous unlabelled PtdInsP2 as a substrate. The product was measured by isotope-dilution assay and identified as authentic Ins(1,4,5)P3. Since phospholipase C is strictly Ca(2+)-dependent, with an optimal concentration range of 1-100 microM, cell lysates were prepared in EGTA and the enzyme reaction was started by adding 10 microM free Ca2+. Phospholipase C activity increased 2-fold during Dictyostelium development up to 8 h of starvation, after which the activity declined to less than 10% of the vegetative level. Enzyme activity in vitro increased up to 2-fold after stimulation of cells with the agonist cAMP in vivo. Addition of 10 microM guanosine 5'-[gamma-thio]triphosphate during lysis activated the enzyme to the same extent, and this effect was antagonized by guanosine 5'-[beta-thio]diphosphate. These results strongly suggest that surface cAMP receptors and G-proteins regulate phospholipase C during Dictyostelium development.

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Selected References

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

  1. Ashkenazi A., Peralta E. G., Winslow J. W., Ramachandran J., Capon D. J. Functionally distinct G proteins selectively couple different receptors to PI hydrolysis in the same cell. Cell. 1989 Feb 10;56(3):487–493. doi: 10.1016/0092-8674(89)90251-1. [DOI] [PubMed] [Google Scholar]
  2. Baldassare J. J., Henderson P. A., Fisher G. J. Isolation and characterization of one soluble and two membrane-associated forms of phosphoinositide-specific phospholipase C from human platelets. Biochemistry. 1989 Jul 11;28(14):6010–6016. doi: 10.1021/bi00440a043. [DOI] [PubMed] [Google Scholar]
  3. Banno Y., Yu A., Nakashima T., Homma Y., Takenawa T., Nozawa Y. Purification and characterization of a cytosolic phosphoinositide-phospholipase C (gamma 2-type) from human platelets. Biochem Biophys Res Commun. 1990 Mar 16;167(2):396–401. doi: 10.1016/0006-291x(90)92035-x. [DOI] [PubMed] [Google Scholar]
  4. Berridge M. J., Irvine R. F. Inositol phosphates and cell signalling. Nature. 1989 Sep 21;341(6239):197–205. doi: 10.1038/341197a0. [DOI] [PubMed] [Google Scholar]
  5. Berridge M. J., Irvine R. F. Inositol trisphosphate, a novel second messenger in cellular signal transduction. Nature. 1984 Nov 22;312(5992):315–321. doi: 10.1038/312315a0. [DOI] [PubMed] [Google Scholar]
  6. Bominaar A. A., Van Haastert P. J. Phospholipase C in Dictyostelium discoideum. Identification of stimulatory and inhibitory surface receptors and G-proteins. Biochem J. 1994 Jan 1;297(Pt 1):189–193. doi: 10.1042/bj2970189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Caterina M. J., Devreotes P. N. Molecular insights into eukaryotic chemotaxis. FASEB J. 1991 Dec;5(15):3078–3085. [PubMed] [Google Scholar]
  8. Cubitt A. B., Firtel R. A. Characterization of phospholipase activity in Dictyostelium discoideum. Identification of a Ca(2+)-dependent polyphosphoinositide-specific phospholipase C. Biochem J. 1992 Apr 15;283(Pt 2):371–378. doi: 10.1042/bj2830371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Deckmyn H., Tu S. M., Majerus P. W. Guanine nucleotides stimulate soluble phosphoinositide-specific phospholipase C in the absence of membranes. J Biol Chem. 1986 Dec 15;261(35):16553–16558. [PubMed] [Google Scholar]
  10. Drayer A. L., van Haastert P. J. Molecular cloning and expression of a phosphoinositide-specific phospholipase C of Dictyostelium discoideum. J Biol Chem. 1992 Sep 15;267(26):18387–18392. [PubMed] [Google Scholar]
  11. Emori Y., Homma Y., Sorimachi H., Kawasaki H., Nakanishi O., Suzuki K., Takenawa T. A second type of rat phosphoinositide-specific phospholipase C containing a src-related sequence not essential for phosphoinositide-hydrolyzing activity. J Biol Chem. 1989 Dec 25;264(36):21885–21890. [PubMed] [Google Scholar]
  12. Europe-Finner G. N., Gammon B., Wood C. A., Newell P. C. Inositol tris- and polyphosphate formation during chemotaxis of Dictyostelium. J Cell Sci. 1989 Aug;93(Pt 4):585–592. doi: 10.1242/jcs.93.4.585. [DOI] [PubMed] [Google Scholar]
  13. Europe-Finner G. N., Newell P. C. GTP analogues stimulate inositol trisphosphate formation transiently in Dictyostelium. J Cell Sci. 1987 May;87(Pt 4):513–518. doi: 10.1242/jcs.87.4.513. [DOI] [PubMed] [Google Scholar]
  14. Europe-Finner G. N., Newell P. C. Inositol 1,4,5-triphosphate induces calcium release from a non- mitochondrial pool in amoebae of Dictyostelium. Biochim Biophys Acta. 1986 Aug 1;887(3):335–340. doi: 10.1016/0167-4889(86)90163-1. [DOI] [PubMed] [Google Scholar]
  15. Fukui T., Lutz R. J., Lowenstein J. M. Purification of a phospholipase C from rat liver cytosol that acts on phosphatidylinositol 4,5-bisphosphate and phosphatidylinositol 4-phosphate. J Biol Chem. 1988 Nov 25;263(33):17730–17737. [PubMed] [Google Scholar]
  16. Gierschik P., Jakobs K. H. Receptor-mediated ADP-ribosylation of a phospholipase C-stimulating G protein. FEBS Lett. 1987 Nov 16;224(1):219–223. doi: 10.1016/0014-5793(87)80451-9. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Kim H. K., Kim J. W., Zilberstein A., Margolis B., Kim J. G., Schlessinger J., Rhee S. G. PDGF stimulation of inositol phospholipid hydrolysis requires PLC-gamma 1 phosphorylation on tyrosine residues 783 and 1254. Cell. 1991 May 3;65(3):435–441. doi: 10.1016/0092-8674(91)90461-7. [DOI] [PubMed] [Google Scholar]
  19. Kishimoto A., Takai Y., Mori T., Kikkawa U., Nishizuka Y. Activation of calcium and phospholipid-dependent protein kinase by diacylglycerol, its possible relation to phosphatidylinositol turnover. J Biol Chem. 1980 Mar 25;255(6):2273–2276. [PubMed] [Google Scholar]
  20. 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]
  21. Lundberg G. A., Newell P. C. Membrane-associated phosphoinositidase C activity in Dictyostelium discoideum. FEBS Lett. 1990 Sep 17;270(1-2):181–183. doi: 10.1016/0014-5793(90)81262-m. [DOI] [PubMed] [Google Scholar]
  22. Meldrum E., Katan M., Parker P. A novel inositol-phospholipid-specific phospholipase C. Rapid purification and characterization. Eur J Biochem. 1989 Jul 1;182(3):673–677. doi: 10.1111/j.1432-1033.1989.tb14878.x. [DOI] [PubMed] [Google Scholar]
  23. Melin P. M., Sundler R., Jergil B. Phospholipase C in rat liver plasma membranes. Phosphoinositide specificity and regulation by guanine nucleotides and calcium. FEBS Lett. 1986 Mar 17;198(1):85–88. doi: 10.1016/0014-5793(86)81189-9. [DOI] [PubMed] [Google Scholar]
  24. Moolenaar W. H., Bierman A. J., Tilly B. C., Verlaan I., Defize L. H., Honegger A. M., Ullrich A., Schlessinger J. A point mutation at the ATP-binding site of the EGF-receptor abolishes signal transduction. EMBO J. 1988 Mar;7(3):707–710. doi: 10.1002/j.1460-2075.1988.tb02866.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nakashima S., Hattori H., Shirato L., Takenaka A., Nozawa Y. Differential sensitivity of arachidonic acid release and 1,2-diacylglycerol formation to pertussis toxin, GDP beta S and NaF in saponin-permeabilized human platelets: possible evidence for distinct GTP-binding proteins involving phospholipase C and A2 activation. Biochem Biophys Res Commun. 1987 Nov 13;148(3):971–978. doi: 10.1016/s0006-291x(87)80227-9. [DOI] [PubMed] [Google Scholar]
  26. Rhee S. G., Suh P. G., Ryu S. H., Lee S. Y. Studies of inositol phospholipid-specific phospholipase C. Science. 1989 May 5;244(4904):546–550. doi: 10.1126/science.2541501. [DOI] [PubMed] [Google Scholar]
  27. Saxe C. L., 3rd, Klein P., Sun T. J., Kimmel A. R., Devreotes P. N. Structure and expression of the cAMP cell-surface receptor. Dev Genet. 1988;9(4-5):227–235. doi: 10.1002/dvg.1020090405. [DOI] [PubMed] [Google Scholar]
  28. Smrcka A. V., Hepler J. R., Brown K. O., Sternweis P. C. Regulation of polyphosphoinositide-specific phospholipase C activity by purified Gq. Science. 1991 Feb 15;251(4995):804–807. doi: 10.1126/science.1846707. [DOI] [PubMed] [Google Scholar]
  29. Taylor S. J., Exton J. H. Guanine-nucleotide and hormone regulation of polyphosphoinositide phospholipase C activity of rat liver plasma membranes. Bivalent-cation and phospholipid requirements. Biochem J. 1987 Dec 15;248(3):791–799. doi: 10.1042/bj2480791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Van Haastert P. J. Determination of inositol 1,4,5-trisphosphate levels in Dictyostelium by isotope dilution assay. Anal Biochem. 1989 Feb 15;177(1):115–119. doi: 10.1016/0003-2697(89)90024-9. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Van Lookeren Campagne M. M., Erneux C., Van Eijk R., Van Haastert P. J. Two dephosphorylation pathways of inositol 1,4,5-trisphosphate in homogenates of the cellular slime mould Dictyostelium discoideum. Biochem J. 1988 Sep 1;254(2):343–350. doi: 10.1042/bj2540343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Weeks G. The manipulation of the fatty acid composition of Dictyostelium discoideum and its effect on cell differentiation. Biochim Biophys Acta. 1976 Oct 21;450(1):21–32. doi: 10.1016/0005-2760(76)90295-2. [DOI] [PubMed] [Google Scholar]
  35. Wilson D. B., Bross T. E., Hofmann S. L., Majerus P. W. Hydrolysis of polyphosphoinositides by purified sheep seminal vesicle phospholipase C enzymes. J Biol Chem. 1984 Oct 10;259(19):11718–11724. [PubMed] [Google Scholar]

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