Skip to main content

Some NLM-NCBI services and products are experiencing heavy traffic, which may affect performance and availability. We apologize for the inconvenience and appreciate your patience. For assistance, please contact our Help Desk at info@ncbi.nlm.nih.gov.

Biochemical Journal logoLink to Biochemical Journal
. 1985 May 1;227(3):971–979. doi: 10.1042/bj2270971

Breakdown of phosphatidylinositol 4,5-bisphosphate in a T-cell leukaemia line stimulated by phytohaemagglutinin is not dependent on Ca2+ mobilization.

T Sasaki, H Hasegawa-Sasaki
PMCID: PMC1144929  PMID: 2988510

Abstract

Addition of phytohaemagglutinin (PHA) to the [32P]Pi-prelabelled JURKAT cells, a human T-cell leukaemia line, resulted in a decrease of [32P]phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] to about 35% of the control value. The decrease was almost complete within 30s after the PHA addition. This decrease was followed by an increase in the 32P-labelling of phosphatidic acid (maximally 2.8-fold at 2 min). The stimulation of myo-[2-3H]inositol-prelabelled JURKAT cells by PHA induced an accumulation of [2-3H]inositol trisphosphate in the presence of 5 mM-LiCl. The result indicates hydrolysis of PtdIns (4,5)P2 by a phospholipase C. The PHA stimulation of JURKAT cells induced about 6-fold increase in the cytosolic free Ca2+ concentration, [Ca2+]i, which was reported by Quin-2, a fluorescent Ca2+ indicator. Studies with partially Ca2+-depleted JURKAT cells, with the Ca2+ ionophore A23187, and with 8-(diethylamino)-octyl-3,4,5-trimethoxybenzoate indicate that the breakdown of PtdIns(4,5)P2 is not mediated through changes of [Ca2+]i. These results therefore indicate that the PHA-induced breakdown of PtdIns(4,5)P2 in JURKAT cells is not dependent on the Ca2+ mobilization.

Full text

PDF
971

Images in this article

Selected References

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

  1. Abdel-Latif A. A., Akhtar R. A., Hawthorne J. N. Acetylcholine increases the breakdown of triphosphoinositide of rabbit iris muscle prelabelled with [32P] phosphate. Biochem J. 1977 Jan 15;162(1):61–73. doi: 10.1042/bj1620061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Agranoff B. W., Murthy P., Seguin E. B. Thrombin-induced phosphodiesteratic cleavage of phosphatidylinositol bisphosphate in human platelets. J Biol Chem. 1983 Feb 25;258(4):2076–2078. [PubMed] [Google Scholar]
  3. Berridge M. J., Dawson R. M., Downes C. P., Heslop J. P., Irvine R. F. Changes in the levels of inositol phosphates after agonist-dependent hydrolysis of membrane phosphoinositides. Biochem J. 1983 May 15;212(2):473–482. doi: 10.1042/bj2120473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Berridge M. J. Inositol trisphosphate and diacylglycerol as second messengers. Biochem J. 1984 Jun 1;220(2):345–360. doi: 10.1042/bj2200345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Berridge M. J. Rapid accumulation of inositol trisphosphate reveals that agonists hydrolyse polyphosphoinositides instead of phosphatidylinositol. Biochem J. 1983 Jun 15;212(3):849–858. doi: 10.1042/bj2120849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Billah M. M., Lapetina E. G. Degradation of phosphatidylinositol-4,5-bisphosphate is insensitive to CA2+ mobilization in stimulated platelets. Biochem Biophys Res Commun. 1982 Nov 16;109(1):217–222. doi: 10.1016/0006-291x(82)91587-x. [DOI] [PubMed] [Google Scholar]
  7. Billah M. M., Lapetina E. G. Platelet-activating factor stimulates metabolism of phosphoinositides in horse platelets: possible relationship to Ca2+ mobilization during stimulation. Proc Natl Acad Sci U S A. 1983 Feb;80(4):965–968. doi: 10.1073/pnas.80.4.965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Billah M. M., Lapetina E. G. Rapid decrease of phosphatidylinositol 4,5-bisphosphate in thrombin-stimulated platelets. J Biol Chem. 1982 Nov 10;257(21):12705–12708. [PubMed] [Google Scholar]
  9. Charest R., Blackmore P. F., Berthon B., Exton J. H. Changes in free cytosolic Ca2+ in hepatocytes following alpha 1-adrenergic stimulation. Studies on Quin-2-loaded hepatocytes. J Biol Chem. 1983 Jul 25;258(14):8769–8773. [PubMed] [Google Scholar]
  10. Chiou C. Y., Malagodi M. H. Studies on the mechanism of action of a new Ca-2+ antagonist, 8-(N,N-diethylamino)octyl 3,4,5-trimethoxybenzoate hydrochloride in smooth and skeletal muscles. Br J Pharmacol. 1975 Feb;53(2):279–285. doi: 10.1111/j.1476-5381.1975.tb07359.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Creba J. A., Downes C. P., Hawkins P. T., Brewster G., Michell R. H., Kirk C. J. Rapid breakdown of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate in rat hepatocytes stimulated by vasopressin and other Ca2+-mobilizing hormones. Biochem J. 1983 Jun 15;212(3):733–747. doi: 10.1042/bj2120733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Downes C. P., Wusteman M. M. Breakdown of polyphosphoinositides and not phosphatidylinositol accounts for muscarinic agonist-stimulated inositol phospholipid metabolism in rat parotid glands. Biochem J. 1983 Dec 15;216(3):633–640. doi: 10.1042/bj2160633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ferber E., Reilly C. E., Resch K. Phospholipid metabolism of stimulated lymphocytes. Comparison of the activation of acyl-CoA:lysolecithin acyltransferase with the binding of concanavalin A to thymocytes. Biochim Biophys Acta. 1976 Sep 21;448(1):143–154. doi: 10.1016/0005-2736(76)90083-3. [DOI] [PubMed] [Google Scholar]
  14. Fisher D. B., Mueller G. C. An early alteration in the phospholipid metabolism of lymphocytes by phytohemagglutinin. Proc Natl Acad Sci U S A. 1968 Aug;60(4):1396–1402. doi: 10.1073/pnas.60.4.1396. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gillis S., Watson J. Biochemical and biological characterization of lymphocyte regulatory molecules. V. Identification of an interleukin 2-producing human leukemia T cell line. J Exp Med. 1980 Dec 1;152(6):1709–1719. doi: 10.1084/jem.152.6.1709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hallcher L. M., Sherman W. R. The effects of lithium ion and other agents on the activity of myo-inositol-1-phosphatase from bovine brain. J Biol Chem. 1980 Nov 25;255(22):10896–10901. [PubMed] [Google Scholar]
  17. Hasegawa-Sasaki H., Sasaki T. Phytohemagglutinin induces rapid degradation of phosphatidylinositol 4,5-bisphosphate and transient accumulation of phosphatidic acid and diacylglycerol in a human T lymphoblastoid cell line, CCRF-CEM. Biochim Biophys Acta. 1983 Dec 20;754(3):305–314. [PubMed] [Google Scholar]
  18. Hasegawa-Sasaki H., Sasaki T. Phytomitogen-induced stimulation of syntheses de novo of phosphatidylinositol, phosphatidic acid and diacylglycerol in rat and human lymphocytes. Biochim Biophys Acta. 1981 Nov 23;666(2):252–258. [PubMed] [Google Scholar]
  19. Hasegawa-Sasaki H., Sasaki T. Rapid breakdown of phosphatidylinositol accompanied by accumulation of phosphatidic acid and diacylglycerol in rat lymphocytes stimulated by concanavalin A. J Biochem. 1982 Feb;91(2):463–468. doi: 10.1093/oxfordjournals.jbchem.a133718. [DOI] [PubMed] [Google Scholar]
  20. Hesketh T. R., Smith G. A., Moore J. P., Taylor M. V., Metcalfe J. C. Free cytoplasmic calcium concentration and the mitogenic stimulation of lymphocytes. J Biol Chem. 1983 Apr 25;258(8):4876–4882. [PubMed] [Google Scholar]
  21. Joseph S. K., Thomas A. P., Williams R. J., Irvine R. F., Williamson J. R. myo-Inositol 1,4,5-trisphosphate. A second messenger for the hormonal mobilization of intracellular Ca2+ in liver. J Biol Chem. 1984 Mar 10;259(5):3077–3081. [PubMed] [Google Scholar]
  22. KREBS H. A. Body size and tissue respiration. Biochim Biophys Acta. 1950 Jan;4(1-3):249–269. doi: 10.1016/0006-3002(50)90032-1. [DOI] [PubMed] [Google Scholar]
  23. Kirk C. J., Creba J. A., Downes C. P., Michell R. H. Hormone-stimulated metabolism of inositol lipids and its relationship to hepatic receptor function. Biochem Soc Trans. 1981 Oct;9(5):377–379. doi: 10.1042/bst0090377. [DOI] [PubMed] [Google Scholar]
  24. Larsson E. L., Coutinho A. The role of mitogenic lectins in T-cell triggering. Nature. 1979 Jul 19;280(5719):239–241. doi: 10.1038/280239a0. [DOI] [PubMed] [Google Scholar]
  25. Maino V. C., Hayman M. J., Crumpton M. J. Relationship between enhanced turnover of phosphatidylinositol and lymphocyte activation by mitogens. Biochem J. 1975 Jan;146(1):247–252. doi: 10.1042/bj1460247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Malagodi M. H., Chiou C. Y. Pharmacological evaluation of a new Ca++ antagonist, 8-(N,N-diethylamino)octyl 3,4,5-trimethoxybenzoate hydrochloride (TMB-8): studies in skeletal muscles. Pharmacology. 1974;12(1):20–31. doi: 10.1159/000136517. [DOI] [PubMed] [Google Scholar]
  27. Martin T. F. Thyrotropin-releasing hormone rapidly activates the phosphodiester hydrolysis of polyphosphoinositides in GH3 pituitary cells. Evidence for the role of a polyphosphoinositide-specific phospholipase C in hormone action. J Biol Chem. 1983 Dec 25;258(24):14816–14822. [PubMed] [Google Scholar]
  28. Masuzawa Y., Osawa T., Inoue K., Nojima S. Effects of various mitogens on the phospholipid metabolism of human peripheral lymphocytes. Biochim Biophys Acta. 1973 Dec 20;326(3):339–344. [PubMed] [Google Scholar]
  29. Matsumoto T., Takeshige K., Minakami S. Inhibition of phagocytotic metabolic changes of leukocytes by an intracellular calcium-antagonist 8-(N,N-diethylamino)-octyl-3,4,5-trimethoxybenzoate. Biochem Biophys Res Commun. 1979 Jun 13;88(3):974–979. doi: 10.1016/0006-291x(79)91503-1. [DOI] [PubMed] [Google Scholar]
  30. Michell R. H. Inositol phospholipids and cell surface receptor function. Biochim Biophys Acta. 1975 Mar 25;415(1):81–47. doi: 10.1016/0304-4157(75)90017-9. [DOI] [PubMed] [Google Scholar]
  31. Palacios R. Mechanism of T cell activation: role and functional relationship of HLA-DR antigens and interleukins. Immunol Rev. 1982;63:73–110. doi: 10.1111/j.1600-065x.1982.tb00412.x. [DOI] [PubMed] [Google Scholar]
  32. Prpić V., Blackmore P. F., Exton J. H. Phosphatidylinositol breakdown induced by vasopressin and epinephrine in hepatocytes is calcium-dependent. J Biol Chem. 1982 Oct 10;257(19):11323–11331. [PubMed] [Google Scholar]
  33. Putney J. W., Jr, Burgess G. M., Halenda S. P., McKinney J. S., Rubin R. P. Effects of secretagogues on [32P]phosphatidylinositol 4,5-bisphosphate metabolism in the exocrine pancreas. Biochem J. 1983 May 15;212(2):483–488. doi: 10.1042/bj2120483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rebecchi M. J., Gershengorn M. C. Thyroliberin stimulates rapid hydrolysis of phosphatidylinositol 4,5-bisphosphate by a phosphodiesterase in rat mammotropic pituitary cells. Evidence for an early Ca2+-independent action. Biochem J. 1983 Nov 15;216(2):287–294. doi: 10.1042/bj2160287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Rhodes D., Prpić V., Exton J. H., Blackmore P. F. Stimulation of phosphatidylinositol 4,5-bisphosphate hydrolysis in hepatocytes by vasopressin. J Biol Chem. 1983 Mar 10;258(5):2770–2773. [PubMed] [Google Scholar]
  36. Rittenhouse-Simmons S., Deykin D. The activation by Ca2+ of platelet phospholipase A2. Effects of dibutyryl cyclic adenosine monophosphate and 8-(N,N-diethylamino)-octyl-3,4,5-trimethoxybenzoate. Biochim Biophys Acta. 1978 Nov 1;543(4):409–422. doi: 10.1016/0304-4165(78)90296-9. [DOI] [PubMed] [Google Scholar]
  37. Robb R. J., Munck A., Smith K. A. T cell growth factor receptors. Quantitation, specificity, and biological relevance. J Exp Med. 1981 Nov 1;154(5):1455–1474. doi: 10.1084/jem.154.5.1455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Schacht J. Inhibition by neomycin of polyphosphoinositide turnover in subcellular fractions of guinea-pig cerebral cortex in vitro. J Neurochem. 1976 Nov;27(5):1119–1124. doi: 10.1111/j.1471-4159.1976.tb00318.x. [DOI] [PubMed] [Google Scholar]
  39. Smith K. A., Ruscetti F. W. T-cell growth factor and the culture of cloned functional T cells. Adv Immunol. 1981;31:137–175. doi: 10.1016/s0065-2776(08)60920-7. [DOI] [PubMed] [Google Scholar]
  40. Streb H., Irvine R. F., Berridge M. J., Schulz I. Release of Ca2+ from a nonmitochondrial intracellular store in pancreatic acinar cells by inositol-1,4,5-trisphosphate. Nature. 1983 Nov 3;306(5938):67–69. doi: 10.1038/306067a0. [DOI] [PubMed] [Google Scholar]
  41. Thomas A. P., Marks J. S., Coll K. E., Williamson J. R. Quantitation and early kinetics of inositol lipid changes induced by vasopressin in isolated and cultured hepatocytes. J Biol Chem. 1983 May 10;258(9):5716–5725. [PubMed] [Google Scholar]
  42. Tsien R. Y., Pozzan T., Rink T. J. Calcium homeostasis in intact lymphocytes: cytoplasmic free calcium monitored with a new, intracellularly trapped fluorescent indicator. J Cell Biol. 1982 Aug;94(2):325–334. doi: 10.1083/jcb.94.2.325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Tsien R. Y., Pozzan T., Rink T. J. T-cell mitogens cause early changes in cytoplasmic free Ca2+ and membrane potential in lymphocytes. Nature. 1982 Jan 7;295(5844):68–71. doi: 10.1038/295068a0. [DOI] [PubMed] [Google Scholar]
  44. Vickers J. D., Kinlough-Rathbone R. L., Mustard J. F. Changes in phosphatidylinositol-4,5-bisphosphate 10 seconds after stimulation of washed rabbit platelets with ADP. Blood. 1982 Dec;60(6):1247–1250. [PubMed] [Google Scholar]
  45. Weiss S. J., McKinney J. S., Putney J. W., Jr Receptor-mediated net breakdown of phosphatidylinositol 4,5-bisphosphate in parotid acinar cells. Biochem J. 1982 Sep 15;206(3):555–560. doi: 10.1042/bj2060555. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES