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. 1994 Nov 25;22(23):5093–5098. doi: 10.1093/nar/22.23.5093

Characterization of the promoter for the human 85 kDa cytosolic phospholipase A2 gene.

T Wu 1, T Ikezono 1, C W Angus 1, J H Shelhamer 1
PMCID: PMC523783  PMID: 7800505

Abstract

The 85 kDa cytosolic phospholipase A2 (cPLA2) plays a key role in the production of arachidonic acid and lysophospholipids, the precursors of eicosanoids and platelet-activating factor. Here we report the cloning of the promoter of the human cPLA2 gene. A 5.7 kb EcoRI fragment containing the most 5' region of the cPLA2 cDNA was sequenced. The transcription initiation site was identified by rapid amplification of 5'-cDNA ends (5'-RACE) and primer extension analysis. DNA sequence analysis of the 595 base pairs 5' of the transcription start site reveals a 48 base purine-pyrimidine dinucleotide repeat (CA repeat), five interferon-gamma response elements (gamma-IRE), one interferon-gamma activated sequence (GAS) and two glucocorticoid response elements (GRE). The promoter lacks a TATA box. It contains a possible CAAT box at -111 and two octamer binding motifs. The 595 base fragment located immediately upstream of the transcriptional start site exhibited functional promoter activity in transient transfection assays in a bronchial epithelial cell line (BEAS 2B cells). Deletion analysis revealed that the CA repeat may confer an inhibitory effect on the cPLA2 promoter activity. The characterization of the human cPLA2 promoter sequence will allow further studies defining the molecular events regulating the expression of the cPLA2 enzyme, especially the cytokine mediated cPLA2 gene expression.

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Selected References

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  1. Azizkhan J. C., Jensen D. E., Pierce A. J., Wade M. Transcription from TATA-less promoters: dihydrofolate reductase as a model. Crit Rev Eukaryot Gene Expr. 1993;3(4):229–254. [PubMed] [Google Scholar]
  2. Benoist C., O'Hare K., Breathnach R., Chambon P. The ovalbumin gene-sequence of putative control regions. Nucleic Acids Res. 1980 Jan 11;8(1):127–142. doi: 10.1093/nar/8.1.127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. Dennis E. A. Diversity of group types, regulation, and function of phospholipase A2. J Biol Chem. 1994 May 6;269(18):13057–13060. [PubMed] [Google Scholar]
  7. Dennis E. A., Rhee S. G., Billah M. M., Hannun Y. A. Role of phospholipase in generating lipid second messengers in signal transduction. FASEB J. 1991 Apr;5(7):2068–2077. doi: 10.1096/fasebj.5.7.1901288. [DOI] [PubMed] [Google Scholar]
  8. Diez E., Mong S. Purification of a phospholipase A2 from human monocytic leukemic U937 cells. Calcium-dependent activation and membrane association. J Biol Chem. 1990 Aug 25;265(24):14654–14661. [PubMed] [Google Scholar]
  9. Doerfler M. E., Weiss J., Clark J. D., Elsbach P. Bacterial lipopolysaccharide primes human neutrophils for enhanced release of arachidonic acid and causes phosphorylation of an 85-kD cytosolic phospholipase A2. J Clin Invest. 1994 Apr;93(4):1583–1591. doi: 10.1172/JCI117138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Eichbaum Q. G., Iyer R., Raveh D. P., Mathieu C., Ezekowitz R. A. Restriction of interferon gamma responsiveness and basal expression of the myeloid human Fc gamma R1b gene is mediated by a functional PU.1 site and a transcription initiator consensus. J Exp Med. 1994 Jun 1;179(6):1985–1996. doi: 10.1084/jem.179.6.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Faisst S., Meyer S. Compilation of vertebrate-encoded transcription factors. Nucleic Acids Res. 1992 Jan 11;20(1):3–26. doi: 10.1093/nar/20.1.3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Jantzen K., Fritton H. P., Igo-Kemenes T., Espel E., Janich S., Cato A. C., Mugele K., Beato M. Partial overlapping of binding sequences for steroid hormone receptors and DNaseI hypersensitive sites in the rabbit uteroglobin gene region. Nucleic Acids Res. 1987 Jun 11;15(11):4535–4552. doi: 10.1093/nar/15.11.4535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kanno Y., Kozak C. A., Schindler C., Driggers P. H., Ennist D. L., Gleason S. L., Darnell J. E., Jr, Ozato K. The genomic structure of the murine ICSBP gene reveals the presence of the gamma interferon-responsive element, to which an ISGF3 alpha subunit (or similar) molecule binds. Mol Cell Biol. 1993 Jul;13(7):3951–3963. doi: 10.1128/mcb.13.7.3951. [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. Ko H. S., Fast P., McBride W., Staudt L. M. A human protein specific for the immunoglobulin octamer DNA motif contains a functional homeobox domain. Cell. 1988 Oct 7;55(1):135–144. doi: 10.1016/0092-8674(88)90016-5. [DOI] [PubMed] [Google Scholar]
  17. Kramer R. M., Hession C., Johansen B., Hayes G., McGray P., Chow E. P., Tizard R., Pepinsky R. B. Structure and properties of a human non-pancreatic phospholipase A2. J Biol Chem. 1989 Apr 5;264(10):5768–5775. [PubMed] [Google Scholar]
  18. 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]
  19. Kramer R. M., Roberts E. F., Manetta J., Putnam J. E. The Ca2(+)-sensitive cytosolic phospholipase A2 is a 100-kDa protein in human monoblast U937 cells. J Biol Chem. 1991 Mar 15;266(8):5268–5272. [PubMed] [Google Scholar]
  20. Lew D. J., Decker T., Strehlow I., Darnell J. E. Overlapping elements in the guanylate-binding protein gene promoter mediate transcriptional induction by alpha and gamma interferons. Mol Cell Biol. 1991 Jan;11(1):182–191. doi: 10.1128/mcb.11.1.182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. 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]
  23. 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]
  24. Liscovitch M., Cantley L. C. Lipid second messengers. Cell. 1994 May 6;77(3):329–334. doi: 10.1016/0092-8674(94)90148-1. [DOI] [PubMed] [Google Scholar]
  25. Mayer R. J., Marshall L. A. New insights on mammalian phospholipase A2(s); comparison of arachidonoyl-selective and -nonselective enzymes. FASEB J. 1993 Feb 1;7(2):339–348. doi: 10.1096/fasebj.7.2.8440410. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Naylor L. H., Clark E. M. d(TG)n.d(CA)n sequences upstream of the rat prolactin gene form Z-DNA and inhibit gene transcription. Nucleic Acids Res. 1990 Mar 25;18(6):1595–1601. doi: 10.1093/nar/18.6.1595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. Pearse R. N., Feinman R., Ravetch J. V. Characterization of the promoter of the human gene encoding the high-affinity IgG receptor: transcriptional induction by gamma-interferon is mediated through common DNA response elements. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11305–11309. doi: 10.1073/pnas.88.24.11305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pearse R. N., Feinman R., Shuai K., Darnell J. E., Jr, Ravetch J. V. Interferon gamma-induced transcription of the high-affinity Fc receptor for IgG requires assembly of a complex that includes the 91-kDa subunit of transcription factor ISGF3. Proc Natl Acad Sci U S A. 1993 May 1;90(9):4314–4318. doi: 10.1073/pnas.90.9.4314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Seilhamer J. J., Pruzanski W., Vadas P., Plant S., Miller J. A., Kloss J., Johnson L. K. Cloning and recombinant expression of phospholipase A2 present in rheumatoid arthritic synovial fluid. J Biol Chem. 1989 Apr 5;264(10):5335–5338. [PubMed] [Google Scholar]
  32. 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]
  33. Smale S. T., Baltimore D. The "initiator" as a transcription control element. Cell. 1989 Apr 7;57(1):103–113. doi: 10.1016/0092-8674(89)90176-1. [DOI] [PubMed] [Google Scholar]
  34. Smale S. T., Schmidt M. C., Berk A. J., Baltimore D. Transcriptional activation by Sp1 as directed through TATA or initiator: specific requirement for mammalian transcription factor IID. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4509–4513. doi: 10.1073/pnas.87.12.4509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tay A., Maxwell P., Li Z., Goldberg H., Skorecki K. Isolation of promoter for cytosolic phospholipase A2 (cPLA2). Biochim Biophys Acta. 1994 Apr 6;1217(3):345–347. doi: 10.1016/0167-4781(94)90299-2. [DOI] [PubMed] [Google Scholar]
  36. Tripathi J., Brahmachari S. K. Distribution of simple repetitive (TG/CA)n and (CT/AG)n sequences in human and rodent genomes. J Biomol Struct Dyn. 1991 Oct;9(2):387–397. doi: 10.1080/07391102.1991.10507919. [DOI] [PubMed] [Google Scholar]
  37. Wang I. M., Mehta V., Cook R. G. Regulation of TL antigen expression. Analysis of the T18d promoter region and responses to IFN-gamma. J Immunol. 1993 Sep 1;151(5):2646–2657. [PubMed] [Google Scholar]
  38. Weis L., Reinberg D. Transcription by RNA polymerase II: initiator-directed formation of transcription-competent complexes. FASEB J. 1992 Nov;6(14):3300–3309. doi: 10.1096/fasebj.6.14.1426767. [DOI] [PubMed] [Google Scholar]
  39. Winitz S., Gupta S. K., Qian N. X., Heasley L. E., Nemenoff R. A., Johnson G. L. Expression of a mutant Gi2 alpha subunit inhibits ATP and thrombin stimulation of cytoplasmic phospholipase A2-mediated arachidonic acid release independent of Ca2+ and mitogen-activated protein kinase regulation. J Biol Chem. 1994 Jan 21;269(3):1889–1895. [PubMed] [Google Scholar]
  40. Wu T., Levine S. J., Lawrence M. G., Logun C., Angus C. W., Shelhamer J. H. Interferon-gamma induces the synthesis and activation of cytosolic phospholipase A2. J Clin Invest. 1994 Feb;93(2):571–577. doi: 10.1172/JCI117009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Xing M., Mattera R. Phosphorylation-dependent regulation of phospholipase A2 by G-proteins and Ca2+ in HL60 granulocytes. J Biol Chem. 1992 Dec 25;267(36):25966–25975. [PubMed] [Google Scholar]
  42. Yang Z., Sugawara M., Ponath P. D., Wessendorf L., Banerji J., Li Y., Strominger J. L. Interferon gamma response region in the promoter of the human DPA gene. Proc Natl Acad Sci U S A. 1990 Dec;87(23):9226–9230. doi: 10.1073/pnas.87.23.9226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Zahedi K., Prada A. E., Davis A. E., 3rd Transcriptional regulation of the C1 inhibitor gene by gamma-interferon. J Biol Chem. 1994 Apr 1;269(13):9669–9674. [PubMed] [Google Scholar]

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