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. 1994 Feb;93(2):571–577. doi: 10.1172/JCI117009

Interferon-gamma induces the synthesis and activation of cytosolic phospholipase A2.

T Wu 1, S J Levine 1, M G Lawrence 1, C Logun 1, C W Angus 1, J H Shelhamer 1
PMCID: PMC293880  PMID: 8113394

Abstract

Both IFN-alpha/beta and IFN-gamma have recently been demonstrated to induce a rapid but transient activation of phospholipase A2 (PLA2) in BALB/c 3T3 fibroblasts and a human neuroblastoma cell line. We report that IFN-gamma induces the synthesis and prolonged activation of cytosolic phospholipase A2 (cPLA2) in a human bronchial epithelial cell line (BEAS 2B). Treatment of the cells with IFN-gamma (300 U/ml) increased the release of [3H]arachidonic acid (AA) from prelabeled cells with a maximal effect at 12 h after stimulation. The increased [3H]AA release was inhibited by the PLA2 inhibitor p-bromophenacyl bromide (10(-5) M). Calcium ionophore A23187 (10(-5) M) further increased the [3H]AA release from the IFN-gamma-treated cells. Subcellular enzyme activity assay revealed that IFN-gamma increased PLA2 activity in both the cytosol and membrane fractions with a translocation of the cPLA2 to cell membranes in a Ca(2+)-free cell lysing buffer. Treatment with IFN-gamma also induced the release of 15-HETE, an arachidonic acid metabolite. Immunoblot showed that IFN-gamma induced the synthesis of cPLA2 protein. Nuclear run-on assay demonstrated that IFN-gamma initiated cPLA2 gene transcription within 15 min, and this effect was sustained at 4 h and returned to near control level at 12 h. The cPLA2 mRNA level was assayed by reverse transcription and PCR. IFN-gamma was found to increase the cPLA2 mRNA after 2-24 h treatment. Furthermore, the IFN-gamma induced cPLA2 mRNA increase was blocked by inhibitors of protein kinase C and calcium/calmodulin-dependent protein kinases, suggesting the involvement of these protein kinases in IFN-gamma-induced gene expression of cPLA2. This study shows that IFN-gamma induces the synthesis and prolonged activation of cPLA2.

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Selected References

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

  1. Azzawi M., Johnston P. W., Majumdar S., Kay A. B., Jeffery P. K. T lymphocytes and activated eosinophils in airway mucosa in fatal asthma and cystic fibrosis. Am Rev Respir Dis. 1992 Jun;145(6):1477–1482. doi: 10.1164/ajrccm/145.6.1477. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. 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]
  4. 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]
  5. Conrad D. J., Kuhn H., Mulkins M., Highland E., Sigal E. Specific inflammatory cytokines regulate the expression of human monocyte 15-lipoxygenase. Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):217–221. doi: 10.1073/pnas.89.1.217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Felder C. C., Williams H. L., Axelrod J. A transduction pathway associated with receptors coupled to the inhibitory guanine nucleotide binding protein Gi that amplifies ATP-mediated arachidonic acid release. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6477–6480. doi: 10.1073/pnas.88.15.6477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fu X. Y. A transcription factor with SH2 and SH3 domains is directly activated by an interferon alpha-induced cytoplasmic protein tyrosine kinase(s). Cell. 1992 Jul 24;70(2):323–335. doi: 10.1016/0092-8674(92)90106-m. [DOI] [PubMed] [Google Scholar]
  8. Greenberg M. E., Hermanowski A. L., Ziff E. B. Effect of protein synthesis inhibitors on growth factor activation of c-fos, c-myc, and actin gene transcription. Mol Cell Biol. 1986 Apr;6(4):1050–1057. doi: 10.1128/mcb.6.4.1050. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Greenberg M. E., Ziff E. B. Stimulation of 3T3 cells induces transcription of the c-fos proto-oncogene. Nature. 1984 Oct 4;311(5985):433–438. doi: 10.1038/311433a0. [DOI] [PubMed] [Google Scholar]
  10. Gronich J. H., Bonventre J. V., Nemenoff R. A. Identification and characterization of a hormonally regulated form of phospholipase A2 in rat renal mesangial cells. J Biol Chem. 1988 Nov 15;263(32):16645–16651. [PubMed] [Google Scholar]
  11. Hamid Q., Azzawi M., Ying S., Moqbel R., Wardlaw A. J., Corrigan C. J., Bradley B., Durham S. R., Collins J. V., Jeffery P. K. Expression of mRNA for interleukin-5 in mucosal bronchial biopsies from asthma. J Clin Invest. 1991 May;87(5):1541–1546. doi: 10.1172/JCI115166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hannigan G. E., Williams B. R. Signal transduction by interferon-alpha through arachidonic acid metabolism. Science. 1991 Jan 11;251(4990):204–207. doi: 10.1126/science.1898993. [DOI] [PubMed] [Google Scholar]
  13. Holtzman M. J., Pentland A., Baenziger N. L., Hansbrough J. R. Heterogeneity of cellular expression of arachidonate 15-lipoxygenase: implications for biological activity. Biochim Biophys Acta. 1989 Jun 8;1003(2):204–208. doi: 10.1016/0005-2760(89)90257-9. [DOI] [PubMed] [Google Scholar]
  14. Hunter J. A., Finkbeiner W. E., Nadel J. A., Goetzl E. J., Holtzman M. J. Predominant generation of 15-lipoxygenase metabolites of arachidonic acid by epithelial cells from human trachea. Proc Natl Acad Sci U S A. 1985 Jul;82(14):4633–4637. doi: 10.1073/pnas.82.14.4633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Kavanaugh W. M., Harsh G. R., 4th, Starksen N. F., Rocco C. M., Williams L. T. Transcriptional regulation of the A and B chain genes of platelet-derived growth factor in microvascular endothelial cells. J Biol Chem. 1988 Jun 15;263(17):8470–8472. [PubMed] [Google Scholar]
  17. Lin L. L., Lin A. Y., DeWitt D. L. Interleukin-1 alpha induces the accumulation of cytosolic phospholipase A2 and the release of prostaglandin E2 in human fibroblasts. J Biol Chem. 1992 Nov 25;267(33):23451–23454. [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. McFadden E. R., Jr, Gilbert I. A. Asthma. N Engl J Med. 1992 Dec 31;327(27):1928–1937. doi: 10.1056/NEJM199212313272708. [DOI] [PubMed] [Google Scholar]
  21. Nakamura T., Lin L. L., Kharbanda S., Knopf J., Kufe D. Macrophage colony stimulating factor activates phosphatidylcholine hydrolysis by cytoplasmic phospholipase A2. EMBO J. 1992 Dec;11(13):4917–4922. doi: 10.1002/j.1460-2075.1992.tb05598.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nemenoff R. A., Winitz S., Qian N. X., Van Putten V., Johnson G. L., Heasley L. E. Phosphorylation and activation of a high molecular weight form of phospholipase A2 by p42 microtubule-associated protein 2 kinase and protein kinase C. J Biol Chem. 1993 Jan 25;268(3):1960–1964. [PubMed] [Google Scholar]
  23. Ohashi Y., Motojima S., Fukuda T., Makino S. Airway hyperresponsiveness, increased intracellular spaces of bronchial epithelium, and increased infiltration of eosinophils and lymphocytes in bronchial mucosa in asthma. Am Rev Respir Dis. 1992 Jun;145(6):1469–1476. doi: 10.1164/ajrccm/145.6.1469. [DOI] [PubMed] [Google Scholar]
  24. Ponzoni M., Montaldo P. G., Cornaglia-Ferraris P. Stimulation of receptor-coupled phospholipase A2 by interferon-gamma. FEBS Lett. 1992 Sep 21;310(1):17–21. doi: 10.1016/0014-5793(92)81136-a. [DOI] [PubMed] [Google Scholar]
  25. Reddel R. R., Ke Y., Gerwin B. I., McMenamin M. G., Lechner J. F., Su R. T., Brash D. E., Park J. B., Rhim J. S., Harris C. C. Transformation of human bronchial epithelial cells by infection with SV40 or adenovirus-12 SV40 hybrid virus, or transfection via strontium phosphate coprecipitation with a plasmid containing SV40 early region genes. Cancer Res. 1988 Apr 1;48(7):1904–1909. [PubMed] [Google Scholar]
  26. Rollins B. J., Stiles C. D. Serum-inducible genes. Adv Cancer Res. 1989;53:1–32. doi: 10.1016/s0065-230x(08)60277-8. [DOI] [PubMed] [Google Scholar]
  27. Schalkwijk C. G., Märki F., Van den Bosch H. Studies on the acyl-chain selectivity of cellular phospholipases A2. Biochim Biophys Acta. 1990 May 1;1044(1):139–146. doi: 10.1016/0005-2760(90)90229-q. [DOI] [PubMed] [Google Scholar]
  28. Schindler C., Shuai K., Prezioso V. R., Darnell J. E., Jr Interferon-dependent tyrosine phosphorylation of a latent cytoplasmic transcription factor. Science. 1992 Aug 7;257(5071):809–813. doi: 10.1126/science.1496401. [DOI] [PubMed] [Google Scholar]
  29. Sen G. C., Lengyel P. The interferon system. A bird's eye view of its biochemistry. J Biol Chem. 1992 Mar 15;267(8):5017–5020. [PubMed] [Google Scholar]
  30. Sharp J. D., White D. L., Chiou X. G., Goodson T., Gamboa G. C., McClure D., Burgett S., Hoskins J., Skatrud P. L., Sportsman J. R. Molecular cloning and expression of human Ca(2+)-sensitive cytosolic phospholipase A2. J Biol Chem. 1991 Aug 15;266(23):14850–14853. [PubMed] [Google Scholar]
  31. Spaargaren M., Wissink S., Defize L. H., de Laat S. W., Boonstra J. Characterization and identification of an epidermal-growth-factor-activated phospholipase A2. Biochem J. 1992 Oct 1;287(Pt 1):37–43. doi: 10.1042/bj2870037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tiwari R. K., Kusari J., Kumar R., Sen G. C. Gene induction by interferons and double-stranded RNA: selective inhibition by 2-aminopurine. Mol Cell Biol. 1988 Oct;8(10):4289–4294. doi: 10.1128/mcb.8.10.4289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Velazquez L., Fellous M., Stark G. R., Pellegrini S. A protein tyrosine kinase in the interferon alpha/beta signaling pathway. Cell. 1992 Jul 24;70(2):313–322. doi: 10.1016/0092-8674(92)90105-l. [DOI] [PubMed] [Google Scholar]
  34. Wu T., Lundgren J. D., Rieves R. D., Doerfler M. E., Logun C., Shelhamer J. H. Platelet-activating factor stimulates eicosanoid production in cultured feline tracheal explants. Exp Lung Res. 1991 Nov-Dec;17(6):1079–1094. doi: 10.3109/01902149109064336. [DOI] [PubMed] [Google Scholar]
  35. Wu T., Mullol J., Rieves R. D., Logun C., Hausfield J., Kaliner M. A., Shelhamer J. H. Endothelin-1 stimulates eicosanoid production in cultured human nasal mucosa. Am J Respir Cell Mol Biol. 1992 Feb;6(2):168–174. doi: 10.1165/ajrcmb/6.2.168. [DOI] [PubMed] [Google Scholar]
  36. Wu T., Rieves R. D., Larivee P., Logun C., Lawrence M. G., Shelhamer J. H. Production of eicosanoids in response to endothelin-1 and identification of specific endothelin-1 binding sites in airway epithelial cells. Am J Respir Cell Mol Biol. 1993 Mar;8(3):282–290. doi: 10.1165/ajrcmb/8.3.282. [DOI] [PubMed] [Google Scholar]
  37. Yoshihara Y., Watanabe Y. Translocation of phospholipase A2 from cytosol to membranes in rat brain induced by calcium ions. Biochem Biophys Res Commun. 1990 Jul 31;170(2):484–490. doi: 10.1016/0006-291x(90)92117-i. [DOI] [PubMed] [Google Scholar]
  38. Yu-Lee L. Y., Hrachovy J. A., Stevens A. M., Schwarz L. A. Interferon-regulatory factor 1 is an immediate-early gene under transcriptional regulation by prolactin in Nb2 T cells. Mol Cell Biol. 1990 Jun;10(6):3087–3094. doi: 10.1128/mcb.10.6.3087. [DOI] [PMC free article] [PubMed] [Google Scholar]

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