Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1992 Apr 15;283(Pt 2):371–378. doi: 10.1042/bj2830371

Characterization of phospholipase activity in Dictyostelium discoideum. Identification of a Ca(2+)-dependent polyphosphoinositide-specific phospholipase C.

A B Cubitt 1, R A Firtel 1
PMCID: PMC1131043  PMID: 1315514

Abstract

We have identified a Ca(2+)-dependent polyphosphoinositide-specific phospholipase C activity in Dictyostelium discoideum. Addition of Ca2+ (20 microM) results in the rapid formation of Ins(1,4,5)P3 within 5 s and leads to sustained inositol phosphate production for up to 40 min in membranes prepared from [3H]inositol-labelled cells. The phospholipase C activity is primarily membrane-bound under the conditions used to lyse the cells. In addition to this activity we also identified a family of Ca(2+)-regulated phospholipase activities active on a range of phospholipid substrates, using [3H]palmitate labelling. Inositol-specific phospholipase C activity is highest in vegetatively growing cells and in starved cells during the first 6 h in development, during which time Ca2+ elicited a 5-fold stimulation of inositol phosphate formation. After this time, total activity decreased progressively until 15 h, after which the activity remained constant up until 24 h. During this period, Ca2+ was able to stimulate a 2-fold increase in inositol phosphates.

Full text

PDF
371

Selected References

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

  1. Abe T., Maeda Y., Iijima T. Transient increase of the intracellular Ca2+ concentration during chemotactic signal transduction in Dictyostelium discoideum cells. Differentiation. 1988 Dec;39(2):90–96. doi: 10.1111/j.1432-0436.1988.tb00085.x. [DOI] [PubMed] [Google Scholar]
  2. Bominaar A. A., Van Dijken P., Draijer R., Van Haastert P. J. Developmental regulation of the inositol 1,4,5-trisphosphate phosphatases in Dictyostelium discoideum. Differentiation. 1991 Feb;46(1):1–5. doi: 10.1111/j.1432-0436.1991.tb00859.x. [DOI] [PubMed] [Google Scholar]
  3. Bominaar A. A., van der Kaay J., van Haastert P. J. Dynamics and function of the inositolcycle in Dictyostelium discoideum. Dev Genet. 1991;12(1-2):19–24. doi: 10.1002/dvg.1020120106. [DOI] [PubMed] [Google Scholar]
  4. Boyer J. L., Waldo G. L., Evans T., Northup J. K., Downes C. P., Harden T. K. Modification of AlF-4- and receptor-stimulated phospholipase C activity by G-protein beta gamma subunits. J Biol Chem. 1989 Aug 15;264(23):13917–13922. [PubMed] [Google Scholar]
  5. Carter H. R., Wallace M. A., Fain J. N. Purification and characterization of PLC-beta m, a muscarinic cholinergic regulated phospholipase C from rabbit brain membrane. Biochim Biophys Acta. 1990 Aug 13;1054(1):119–128. doi: 10.1016/0167-4889(90)90213-w. [DOI] [PubMed] [Google Scholar]
  6. DAVIDOFF F., KORN E. D. FATTY ACID AND PHOSPHOLIPID COMPOSITION OF THE CELLULAR SLIME MOLD, DICTYOSTELIUM DISCOIDEUM. THE OCCURRENCE OF PREVIOUSLY UNDESCRIBED FATTY ACIDS. J Biol Chem. 1963 Oct;238:3199–3209. [PubMed] [Google Scholar]
  7. DAVIDOFF F., KORN E. D. THE BIOSYNTHESIS OF FATTY ACIDS IN THE CELLULAR SLIME MOLD, DICTYOSTELIUM DISCOIDEUM. J Biol Chem. 1963 Oct;238:3210–3215. [PubMed] [Google Scholar]
  8. Diaz-Laviada I., Larrodera P., Nieto J. L., Cornet M. E., Diaz-Meco M. T., Sanchez M. J., Guddal P. H., Johansen T., Haro A., Moscat J. Mechanism of inhibition of adenylate cyclase by phospholipase C-catalyzed hydrolysis of phosphatidylcholine. Involvement of a pertussis toxin-sensitive G protein and protein kinase C. J Biol Chem. 1991 Jan 15;266(2):1170–1176. [PubMed] [Google Scholar]
  9. Ellingson J. S. Changes in the phospholipid composition in the differentiating cellular slime mold, Dictyostelium discoideum. Biochim Biophys Acta. 1974 Jan 23;337(1):60–67. doi: 10.1016/0005-2760(74)90040-x. [DOI] [PubMed] [Google Scholar]
  10. Europe-Finner G. N., Ludérus M. E., Small N. V., Van Driel R., Reymond C. D., Firtel R. A., Newell P. C. Mutant ras gene induces elevated levels of inositol tris- and hexakisphosphates in Dictyostelium. J Cell Sci. 1988 Jan;89(Pt 1):13–20. doi: 10.1242/jcs.89.1.13. [DOI] [PubMed] [Google Scholar]
  11. Exton J. H. Signaling through phosphatidylcholine breakdown. J Biol Chem. 1990 Jan 5;265(1):1–4. [PubMed] [Google Scholar]
  12. Ferber E., Munder P. G., Fischer H., Gerisch G. High phospholipase activities in amoebae of Dictyostelium discoideum. Eur J Biochem. 1970 Jun;14(2):253–257. doi: 10.1111/j.1432-1033.1970.tb00284.x. [DOI] [PubMed] [Google Scholar]
  13. Firtel R. A. Signal transduction pathways controlling multicellular development in Dictyostelium. Trends Genet. 1991 Nov-Dec;7(11-12):381–388. doi: 10.1016/0168-9525(91)90260-w. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Fournié J. J., Mullins R. J., Basten A. Isolation and structural characteristics of a monoclonal antibody-defined cross-reactive phospholipid antigen from Mycobacterium tuberculosis and Mycobacterium leprae. J Biol Chem. 1991 Jan 15;266(2):1211–1219. [PubMed] [Google Scholar]
  16. 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]
  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. Hadwiger J. A., Wilkie T. M., Strathmann M., Firtel R. A. Identification of Dictyostelium G alpha genes expressed during multicellular development. Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):8213–8217. doi: 10.1073/pnas.88.18.8213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hase A. Changes in phospholipid composition during the development of Dictyostelium discoideum. Arch Biochem Biophys. 1982 Nov;219(1):21–29. doi: 10.1016/0003-9861(82)90129-1. [DOI] [PubMed] [Google Scholar]
  20. Howard P. K., Ahern K. G., Firtel R. A. Establishment of a transient expression system for Dictyostelium discoideum. Nucleic Acids Res. 1988 Mar 25;16(6):2613–2623. doi: 10.1093/nar/16.6.2613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Imai A., Gershengorn M. C. Measurement of lipid turnover in response to thyrotropin-releasing hormone. Methods Enzymol. 1987;141:100–101. doi: 10.1016/0076-6879(87)41059-8. [DOI] [PubMed] [Google Scholar]
  22. Imai A., Gershengorn M. C. Phosphatidylinositol 4,5-bisphosphate turnover is transient while phosphatidylinositol turnover is persistent in thyrotropin-releasing hormone-stimulated rat pituitary cells. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8540–8544. doi: 10.1073/pnas.83.22.8540. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Janssens P. M., De Jong C. C., Vink A. A., Van Haastert P. J. Regulatory properties of magnesium-dependent guanylate cyclase in Dictyostelium discoideum membranes. J Biol Chem. 1989 Mar 15;264(8):4329–4335. [PubMed] [Google Scholar]
  24. Johnson R. L., Gundersen R., Lilly P., Pitt G. S., Pupillo M., Sun T. J., Vaughan R. A., Devreotes P. N. G-protein-linked signal transduction systems control development in Dictyostelium. Development. 1989;107 (Suppl):75–80. doi: 10.1242/dev.107.Supplement.75. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Kimmel A. R., Firtel R. A. cAMP signal transduction pathways regulating development of Dictyostelium discoideum. Curr Opin Genet Dev. 1991 Oct;1(3):383–390. doi: 10.1016/s0959-437x(05)80304-1. [DOI] [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., 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]
  29. Leslie C. C. Kinetic properties of a high molecular mass arachidonoyl-hydrolyzing phospholipase A2 that exhibits lysophospholipase activity. J Biol Chem. 1991 Jun 15;266(17):11366–11371. [PubMed] [Google Scholar]
  30. Long B. H., Coe E. L. Changes in neutral lipid constituents during differentiation of the cellular slime mold, Dictyostelium discoideum. J Biol Chem. 1974 Jan 25;249(2):521–529. [PubMed] [Google Scholar]
  31. 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]
  32. MacDonald J. I., Weeks G. The biosynthesis and turnover of lipid during the differentiation of Dictyostelium discoideum. Biochim Biophys Acta. 1985 May 17;834(3):301–307. doi: 10.1016/0005-2760(85)90002-5. [DOI] [PubMed] [Google Scholar]
  33. Mehdy M. C., Ratner D., Firtel R. A. Induction and modulation of cell-type-specific gene expression in Dictyostelium. Cell. 1983 Mar;32(3):763–771. doi: 10.1016/0092-8674(83)90062-4. [DOI] [PubMed] [Google Scholar]
  34. Milne J. L., Coukell M. B. A Ca2+ transport system associated with the plasma membrane of Dictyostelium discoideum is activated by different chemoattractant receptors. J Cell Biol. 1991 Jan;112(1):103–110. doi: 10.1083/jcb.112.1.103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Newell P. C., Europe-Finner G. N., Liu G., Gammon B., Wood C. A. Signal transduction for chemotaxis in Dictyostelium amoebae. Semin Cell Biol. 1990 Apr;1(2):105–113. [PubMed] [Google Scholar]
  36. Pitt G. S., Gundersen R. E., Lilly P. J., Pupillo M. B., Vaughan R. A., Devreotes P. N. G protein-linked signal transduction in aggregating Dictyostelium. Soc Gen Physiol Ser. 1990;45:125–131. [PubMed] [Google Scholar]
  37. 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]
  38. 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]
  39. Stephens L. R., Irvine R. F. Stepwise phosphorylation of myo-inositol leading to myo-inositol hexakisphosphate in Dictyostelium. Nature. 1990 Aug 9;346(6284):580–583. doi: 10.1038/346580a0. [DOI] [PubMed] [Google Scholar]
  40. Stephens L. R., Kay R. R., Irvine R. F. A myo-inositol D-3 hydroxykinase activity in Dictyostelium. Biochem J. 1990 Nov 15;272(1):201–210. doi: 10.1042/bj2720201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Stockbridge N. EGTA. Comput Biol Med. 1987;17(5):299–304. doi: 10.1016/0010-4825(87)90019-9. [DOI] [PubMed] [Google Scholar]
  42. 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]
  43. Taylor C. W. The role of G proteins in transmembrane signalling. Biochem J. 1990 Nov 15;272(1):1–13. doi: 10.1042/bj2720001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Theibert A., Palmisano M., Jastorff B., Devreotes P. The specificity of the cAMP receptor mediating activation of adenylate cyclase in Dictyostelium discoideum. Dev Biol. 1986 Apr;114(2):529–533. doi: 10.1016/0012-1606(86)90216-2. [DOI] [PubMed] [Google Scholar]
  45. Tompkins T. A., Moscarello M. A. A 57-kDa phosphatidylinositol-specific phospholipase C from bovine brain. J Biol Chem. 1991 Mar 5;266(7):4228–4236. [PubMed] [Google Scholar]
  46. 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]
  47. Van Haastert P. J. Down-regulation of cell surface cyclic AMP receptors and desensitization of cyclic AMP-stimulated adenylate cyclase by cyclic AMP in Dictyostelium discoideum. Kinetics and concentration dependence. J Biol Chem. 1987 Jun 5;262(16):7700–7704. [PubMed] [Google Scholar]
  48. 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]
  49. 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]
  50. van Blitterswijk W. J., Hilkmann H., de Widt J., van der Bend R. L. Phospholipid metabolism in bradykinin-stimulated human fibroblasts. II. Phosphatidylcholine breakdown by phospholipases C and D; involvement of protein kinase C. J Biol Chem. 1991 Jun 5;266(16):10344–10350. [PubMed] [Google Scholar]
  51. van Haastert P. J., de Wit R. J., Janssens P. M., Kesbeke F., DeGoede J. G-protein-mediated interconversions of cell-surface cAMP receptors and their involvement in excitation and desensitization of guanylate cyclase in Dictyostelium discoideum. J Biol Chem. 1986 May 25;261(15):6904–6911. [PubMed] [Google Scholar]

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

RESOURCES