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. 1995 Jul 1;309(Pt 1):91–97. doi: 10.1042/bj3090091

Arachidonic acid mobilization is suppressed during mitosis: role of cytosolic phospholipase A2 activation.

R D Berlin 1, S F Preston 1
PMCID: PMC1135804  PMID: 7542448

Abstract

In interphase HeLa cells, incubation with histamine or thapsigargin led to the rapid release of arachidonic acid. The release was absolutely dependent on Ca2+, consistent with the activation of an 85 kDa cytosolic phospholipase A2 (cPLA2). In metaphase-arrested HeLa cells, by contrast, the stimulation of arachidonate release by these agents was inhibited by more than 90%. The lack of arachidonic acid release by mitotic cells was at least partly expected, since histamine- or thapsigargin-induced Ca2+ influx and elevations of cytosolic free Ca2+ are known to be strongly inhibited during mitosis [Preston, Sha'afi and Berlin (1991) Cell Regul. 2, 915-925]. Indeed, incubation of interphase cells with the Ca2+ ionophore A23187 alone induced a high level of arachidonate release. However, even A23187 failed to elicit release from mitotic cells. Since the Ca(2+)-dependent release of arachidonate by many cell types is promoted by preincubation with ligands that activate receptors of the tyrosine kinase class, and tumour promoters that lead to the phosphorylation of cPLA2, we determined if the responses of mitotic HeLa cells could be modified by this 'priming' process. We first established that epidermal growth factor and phorbol 12-myristate 13-acetate were effective priming agents in interphase cells: cells preincubated with the hormone or tumour promoter showed a 2-fold stimulation of thapsigargin- or A23187-induced arachidonic acid release. However, none of the priming agents reversed the lack of mitotic cell response. This refractoriness was not caused by destruction of cPLA2 during mitosis: by Western blotting, cPLA2 of interphase and mitotic cells was shown to be present in comparable amounts. Moreover, cPLA2 activities measured in extracts of interphase and mitotic cells were also comparable. Surprisingly, mitotic cPLA2 appeared to be constitutively phosphorylated in non-hormone-treated (control) cells. The results indicate a novel mechanism of regulation by cPLA2 activity in mitotic cells.

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Selected References

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  1. Ackermann E. J., Kempner E. S., Dennis E. A. Ca(2+)-independent cytosolic phospholipase A2 from macrophage-like P388D1 cells. Isolation and characterization. J Biol Chem. 1994 Mar 25;269(12):9227–9233. [PubMed] [Google Scholar]
  2. Berlin R. D., Oliver J. M., Walter R. J. Surface functions during Mitosis I: phagocytosis, pinocytosis and mobility of surface-bound Con A. Cell. 1978 Oct;15(2):327–341. doi: 10.1016/0092-8674(78)90002-8. [DOI] [PubMed] [Google Scholar]
  3. Berlin R. D., Preston S. F. Okadaic acid uncouples calcium entry from depletion of intracellular stores. Cell Calcium. 1993 May;14(5):379–386. doi: 10.1016/0143-4160(93)90042-5. [DOI] [PubMed] [Google Scholar]
  4. Bonney R. C., Beesley J. S., Franks S. Release of arachidonic acid from human endometrial cells in culture mediated by calcium ionophore (A23187) or fluoride. J Reprod Fertil. 1991 Nov;93(2):449–460. doi: 10.1530/jrf.0.0930449. [DOI] [PubMed] [Google Scholar]
  5. Bonventre J. V., Gronich J. H., Nemenoff R. A. Epidermal growth factor enhances glomerular mesangial cell soluble phospholipase A2 activity. J Biol Chem. 1990 Mar 25;265(9):4934–4938. [PubMed] [Google Scholar]
  6. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  7. Brockman J. L., Gross S. D., Sussman M. R., Anderson R. A. Cell cycle-dependent localization of casein kinase I to mitotic spindles. Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9454–9458. doi: 10.1073/pnas.89.20.9454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Channon J. Y., Leslie C. C. A calcium-dependent mechanism for associating a soluble arachidonoyl-hydrolyzing phospholipase A2 with membrane in the macrophage cell line RAW 264.7. J Biol Chem. 1990 Apr 5;265(10):5409–5413. [PubMed] [Google Scholar]
  9. Clark J. D., Lin L. L., Kriz R. W., Ramesha C. S., Sultzman L. A., Lin A. Y., Milona N., Knopf J. L. A novel arachidonic acid-selective cytosolic PLA2 contains a Ca(2+)-dependent translocation domain with homology to PKC and GAP. Cell. 1991 Jun 14;65(6):1043–1051. doi: 10.1016/0092-8674(91)90556-e. [DOI] [PubMed] [Google Scholar]
  10. Clark J. D., Milona N., Knopf J. L. Purification of a 110-kilodalton cytosolic phospholipase A2 from the human monocytic cell line U937. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7708–7712. doi: 10.1073/pnas.87.19.7708. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Durstin M., Durstin S., Molski T. F., Becker E. L., Sha'afi R. I. Cytoplasmic phospholipase A2 translocates to membrane fraction in human neutrophils activated by stimuli that phosphorylate mitogen-activated protein kinase. Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3142–3146. doi: 10.1073/pnas.91.8.3142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Espe U., Fürstenberger G., Marks F., Kaszkin M., Kinzel V. Early changes in the arachidonic acid metabolism of HeLa cells in response to the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) and related compounds. J Cancer Res Clin Oncol. 1987;113(2):137–144. doi: 10.1007/BF00391435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Glaser K. B., Mobilio D., Chang J. Y., Senko N. Phospholipase A2 enzymes: regulation and inhibition. Trends Pharmacol Sci. 1993 Mar;14(3):92–98. doi: 10.1016/0165-6147(93)90071-q. [DOI] [PubMed] [Google Scholar]
  14. Goldberg H. J., Viegas M. M., Margolis B. L., Schlessinger J., Skorecki K. L. The tyrosine kinase activity of the epidermal-growth-factor receptor is necessary for phospholipase A2 activation. Biochem J. 1990 Apr 15;267(2):461–465. doi: 10.1042/bj2670461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  16. Hack N., Margolis B. L., Ullrich A., Schlessinger J., Skorecki K. L. Distinct structural specificities for functional coupling of the epidermal growth factor receptor to calcium-signalling versus phospholipase A2 responses. Biochem J. 1991 May 1;275(Pt 3):563–567. doi: 10.1042/bj2750563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Heider H., Hug C., Lucocq J. M. A 40-kDa myelin basic protein kinase, distinct from erk1 and erk2, is activated in mitotic HeLa cells. Eur J Biochem. 1994 Jan 15;219(1-2):513–520. doi: 10.1111/j.1432-1033.1994.tb19966.x. [DOI] [PubMed] [Google Scholar]
  18. Hesketh T. R., Beaven M. A., Rogers J., Burke B., Warren G. B. Stimulated release of histamine by a rat mast cell line is inhibited during mitosis. J Cell Biol. 1984 Jun;98(6):2250–2254. doi: 10.1083/jcb.98.6.2250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hoeck W. G., Ramesha C. S., Chang D. J., Fan N., Heller R. A. Cytoplasmic phospholipase A2 activity and gene expression are stimulated by tumor necrosis factor: dexamethasone blocks the induced synthesis. Proc Natl Acad Sci U S A. 1993 May 15;90(10):4475–4479. doi: 10.1073/pnas.90.10.4475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jain M. K., Streb M., Rogers J., DeHaas G. H. Action of phospholipase A2 on bilayers containing lysophosphatidylcholine analogs and the effect of inhibitors. Biochem Pharmacol. 1984 Aug 15;33(16):2541–2551. doi: 10.1016/0006-2952(84)90622-1. [DOI] [PubMed] [Google Scholar]
  21. Kast R., Fürstenberger G., Marks F. Activation of cytosolic phospholipase A2 by transforming growth factor-alpha in HEL-30 keratinocytes. J Biol Chem. 1993 Aug 5;268(22):16795–16802. [PubMed] [Google Scholar]
  22. Kramer R. M., Roberts E. F., Manetta J. V., Hyslop P. A., Jakubowski J. A. Thrombin-induced phosphorylation and activation of Ca(2+)-sensitive cytosolic phospholipase A2 in human platelets. J Biol Chem. 1993 Dec 15;268(35):26796–26804. [PubMed] [Google Scholar]
  23. Lin L. L., Lin A. Y., Knopf J. L. Cytosolic phospholipase A2 is coupled to hormonally regulated release of arachidonic acid. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6147–6151. doi: 10.1073/pnas.89.13.6147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lin L. L., Wartmann M., Lin A. Y., Knopf J. L., Seth A., Davis R. J. cPLA2 is phosphorylated and activated by MAP kinase. Cell. 1993 Jan 29;72(2):269–278. doi: 10.1016/0092-8674(93)90666-e. [DOI] [PubMed] [Google Scholar]
  25. Lucocq J. M., Berger E. G., Warren G. Mitotic Golgi fragments in HeLa cells and their role in the reassembly pathway. J Cell Biol. 1989 Aug;109(2):463–474. doi: 10.1083/jcb.109.2.463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Oliver J. M., Seagrave J. C., Pfeiffer J. R., Feibig M. L., Deanin G. G. Surface functions during mitosis in rat basophilic leukemia cells. J Cell Biol. 1985 Dec;101(6):2156–2166. doi: 10.1083/jcb.101.6.2156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Preston S. F., Sha'afi R. I., Berlin R. D. Regulation of Ca2+ influx during mitosis: Ca2+ influx and depletion of intracellular Ca2+ stores are coupled in interphase but not mitosis. Cell Regul. 1991 Nov;2(11):915–925. doi: 10.1091/mbc.2.11.915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Putney J. W., Jr Capacitative calcium entry revisited. Cell Calcium. 1990 Nov-Dec;11(10):611–624. doi: 10.1016/0143-4160(90)90016-n. [DOI] [PubMed] [Google Scholar]
  29. Qiu Z. H., de Carvalho M. S., Leslie C. C. Regulation of phospholipase A2 activation by phosphorylation in mouse peritoneal macrophages. J Biol Chem. 1993 Nov 15;268(32):24506–24513. [PubMed] [Google Scholar]
  30. Ramanadham S., Gross R. W., Han X., Turk J. Inhibition of arachidonate release by secretagogue-stimulated pancreatic islets suppresses both insulin secretion and the rise in beta-cell cytosolic calcium ion concentration. Biochemistry. 1993 Jan 12;32(1):337–346. doi: 10.1021/bi00052a042. [DOI] [PubMed] [Google Scholar]
  31. Sager P. R., Brown P. A., Berlin R. D. Analysis of transferrin recycling in mitotic and interphase HeLa cells by quantitative fluorescence microscopy. Cell. 1984 Dec;39(2 Pt 1):275–282. doi: 10.1016/0092-8674(84)90005-9. [DOI] [PubMed] [Google Scholar]
  32. Samuelsson B., Dahlén S. E., Lindgren J. A., Rouzer C. A., Serhan C. N. Leukotrienes and lipoxins: structures, biosynthesis, and biological effects. Science. 1987 Sep 4;237(4819):1171–1176. doi: 10.1126/science.2820055. [DOI] [PubMed] [Google Scholar]
  33. Tamemoto H., Kadowaki T., Tobe K., Ueki K., Izumi T., Chatani Y., Kohno M., Kasuga M., Yazaki Y., Akanuma Y. Biphasic activation of two mitogen-activated protein kinases during the cell cycle in mammalian cells. J Biol Chem. 1992 Oct 5;267(28):20293–20297. [PubMed] [Google Scholar]
  34. Thastrup O., Cullen P. J., Drøbak B. K., Hanley M. R., Dawson A. P. Thapsigargin, a tumor promoter, discharges intracellular Ca2+ stores by specific inhibition of the endoplasmic reticulum Ca2(+)-ATPase. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2466–2470. doi: 10.1073/pnas.87.7.2466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tsien R. Y. New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures. Biochemistry. 1980 May 27;19(11):2396–2404. doi: 10.1021/bi00552a018. [DOI] [PubMed] [Google Scholar]
  36. Vigo C., Lewis G. P., Piper P. J. Mechanisms of inhibition of phospholipase A2. Biochem Pharmacol. 1980 Feb 15;29(4):623–627. doi: 10.1016/0006-2952(80)90386-x. [DOI] [PubMed] [Google Scholar]
  37. Volpi M., Berlin R. D. Intracellular elevations of free calcium induced by activation of histamine H1 receptors in interphase and mitotic HeLa cells: hormone signal transduction is altered during mitosis. J Cell Biol. 1988 Dec;107(6 Pt 2):2533–2539. doi: 10.1083/jcb.107.6.2533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Warren G., Davoust J., Cockcroft A. Recycling of transferrin receptors in A431 cells is inhibited during mitosis. EMBO J. 1984 Oct;3(10):2217–2225. doi: 10.1002/j.1460-2075.1984.tb02119.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wheatley D. N., Glenesk A., Inglis M. S. Protein synthesis and mitosis. 1. Influence of anti-tubulins on protein synthetic activity in synchronous M-phase and asynchronous populations of HeLa S-3 cells. Cytobios. 1988;55(220):41–50. [PubMed] [Google Scholar]
  40. White S. R., Strek M. E., Kulp G. V., Spaethe S. M., Burch R. A., Neeley S. P., Leff A. R. Regulation of human eosinophil degranulation and activation by endogenous phospholipase A2. J Clin Invest. 1993 May;91(5):2118–2125. doi: 10.1172/JCI116436. [DOI] [PMC free article] [PubMed] [Google Scholar]

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