Skip to main content
SpringerLink
Log in

Activities and interactions among phospholipases A2 during thapsigargin-induced S49 cell death

  • Published:
Apoptosis Aims and scope Submit manuscript

Abstract

The purpose of this study was to determine the roles of calcium-dependent phospholipase A2 (cPLA2) and calcium-independent phospholipase A2 (iPLA2) in thapsigargin-induced membrane susceptibility to secretory phospholipase A2 (sPLA2) and programmed cell death. 3H-arachidonic acid release was observed in the presence of thapsigargin. This release was inhibited partially by an inhibitor of iPLA2 (BEL) and completely by an inhibitor of both cPLA2 and iPLA2 (MAFP) suggesting that these enzymes were active during apoptosis. The process of cell death did not require the activity of either enzyme since neither inhibitor impeded the progression of apoptosis. However, both inhibitors increased the susceptibility of the membrane to sPLA2 in the presence of thapsigargin. In the case of BEL, this effect appeared to involve direct induction of apoptosis in a sub-population of the cells independent of the action of iPLA2. In conclusion, the results suggested that cPLA2 and iPLA2 are active during thapsigargin-induced apoptosis in S49 cells and that cPLA2 tempers the tendency of the cells to become susceptible to sPLA2 during apoptosis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Atsumi G, Murakami M, Tajima M, et al. The perturbed membrane of cells undergoing apoptosis is susceptible to type II secretory phospholipase A2 to liberate arachidonic acid. Biochim Biophys Acta 1997; 1349: 43–54.

    PubMed  Google Scholar 

  2. Nielsen KH, Olsen CA, Allred DV, et al. Susceptibility of S49 lymphoma cell membranes to hydrolysis by secretory phospholipase A2 during early phase of apoptosis. Biochim Biophys Acta 2000; 1484: 163–174.

    PubMed  Google Scholar 

  3. Balsinde J, Dennis EA. Distinct roles in signal transduction for each of the phospholipase enzymes present in P38881 macrophages. J Biol Chem 1996; 271: 6758–6765.

    PubMed  Google Scholar 

  4. Reddy ST, Herschman HR. Prostaglandin synthase-1 and prostaglandin synthase-2 are coupled to distinct phospholipases for the generation of prostaglandin D2 in activated mast cells. J Biol Chem 1997; 272: 3231–3237.

    PubMed  Google Scholar 

  5. Murakami M, Shimbara S, Kambe T, et al. The function of five distinct mammalian phospholipases A2s in regulating arachidonic acid release. J Biol Chem 1998; 273: 14411–14423.

    PubMed  Google Scholar 

  6. Wilson HA, Waldrip JB, Nielson KH, et al. Mechanisms by which elevated intracellular calcium induces S49 cell membranes to become susceptible to the action of secretory phospholipase A2. J Biol Chem 1999; 274: 11494–11504.

    PubMed  Google Scholar 

  7. Lo HH, Teichmann P, Furstenberger G, et al. Suppression or elvation of cytosolic phospholipase A2 alters keratinocyte prostaglandin synthesis, growth and apoptosis. Cancer Res H. A. Wilson et al. 1998; 58: 4624–4631.

    Google Scholar 

  8. Maccarrone M, Nieuwenhuizen WF, Dullens HFJ, et al. Membrane modifications in human erythroleukemia K562 cells during induction of programmed cell death by transforming growth factor beta-1 or cisplatin. Eur J Biochem 1996; 241: 297–302.

    PubMed  Google Scholar 

  9. Wu YL, Jiang XR, Lillington DM, et al. 1,25-Dihydroxyvitamin D-3 protects human leukemic cells from tumor necrosis factor-induced apoptosis via inactivation of cytosolic phospholipase A2. Cancer Res 1998; 58(4): 633–640.

    Google Scholar 

  10. Wu YL, Jiang XR, Newland AC, et al. Failure to activate cytosolic phospholipase A2 causes TNF resistance in human leukemic cells. J Immunol 1998; 160(12): 5929–5935.

    PubMed  Google Scholar 

  11. Adam-Klages S, Schwandner R, Lüschen S, et al. Caspasemediated inhibition of human cytosolic phospholipase A2 during apoptosis. J Immunol 1998; 161: 5687–5694.

    PubMed  Google Scholar 

  12. Dimitrov T, Krajcsi P, Hermiston TW, et al. Adenovirus E3–10.4K/14.5K protein complex inhibits tumor necrosis factorinduced translocation of cytosolic phospholipase A2 to membranes. J Virol 1997; 71: 2830–2837.

    PubMed  Google Scholar 

  13. Fabisiak JP, Kagan VE, Tyurina YY, et al. Paraquat-induced phosphatidylserine oxidation and apoptosis are independent of activation of PLA2. Am J Phys 1998; 18: L793–L802.

    Google Scholar 

  14. Atsumi G, Tajima M, Hadano A, et al. Fas-induced arachidonic acid release is mediated by Ca2+-independent phospholipase A2 but not cytosolic phospholipase A2, which undergoes proteolytic inactivation. J Biol Chem 1998; 273: 13870–13877.

    PubMed  Google Scholar 

  15. Maraganore JM, Merutka G, Cho W, et al. A new class of phspholipases A2 with lysine in place of aspartate 49. J Biol Chem 1984; 259: 13839–13843.

    PubMed  Google Scholar 

  16. Wilson HA, Huang W, Waldrip JB, et al. Mechanisms by which thionin induces susceptibility of S49 cell membrances to extracellular phospholipase A2. Bioch Biophys Acta 1997; 1349: 142–156.

    Google Scholar 

  17. Audette CA, Burnstein SH, Doyle SA, et al. G-protein mediation of cannabinoid-induced phospholipase activation. Pharm Biochem Behav 1991; 40: 559–563.

    Google Scholar 

  18. Bligh EG, Dyer WJ. Can J Biochem Physiol 1959; 37: 911–917.

    PubMed  Google Scholar 

  19. FairbairnDW, O'Neill KL. The neutral comet assay is sufficient to identify an apoptotic ‘window’ by visual inspection. Apoptosis 1996; 1: 91–94.

    Google Scholar 

  20. Cejna M, Fritsch G, Printz D, et al. Kinetics of apoptosis and secondary necrosis in cultured rat thymocytes and S.49 mouse lymphoma and CEM human leukemia cells. Biochem Cell Biol 1994; 72: 677–685.

    PubMed  Google Scholar 

  21. Ackermann EJ, Conde-Frieboes K, Dennis EA. Inhibition of macrophage Ca2+-independent phospholipase A2 by bromoenol lactone and trifluoromethyl ketones. J Biol Chem 1995; 270: 445–450.

    PubMed  Google Scholar 

  22. Lio YC, Reynolds LJ, Balsinde J, et al. Irreversible inhibition of Ca2+-independent phospholipase A2 by methyl arachidonyl fluorophosphonate. Biochim Biophys Acta 1996; 1302(1): 55–60.

    PubMed  Google Scholar 

  23. Lehmen JJ, Brown KA, Ramanadham S, et al. Arachidonic acid release from aortic smooth muscle cells induced by (Arg8)vasopressin is largely mediated by calcium-independent phospholipase A2. J Biol Chem 1993; 268: 20713–20716.

    PubMed  Google Scholar 

  24. Jaattela M, Benedict M, Tewari M, et al. Bcl-x and Bcl-2 inhibit TNF and Fas-induced apoptosis and activation of phospholipase A2 in breast carcinoma cells. Oncogene 1995; 10(12): 2297–2305.

    PubMed  Google Scholar 

  25. Krajcsi P, DimitrovT, Hermiston TW, et al. The adenovirus E3–14.7K protein and the E3–10.4K/14.5K complex of proteins, which independently inhibit tumor necrosis factor (TNF)-induced apoptosis, also independently inhibit TNF-induced release of arachidonic acid. J Virol 1996; 70: 4904–4913.

    PubMed  Google Scholar 

  26. Voelkel-Johnson C, Thorne TE, Laster SM. Susceptibility to TNF in the presence of inhibitors of transcription or translation is dependent on the activity of cytostolic phospholipase A2 in human melanoma tumor cells. J Immunol 1996; 156(1): 201–207.

    PubMed  Google Scholar 

  27. Cai ZZ, Bettaieb A, Elmahdani N, et al. Alteration of the sphingomyelin/ ceramide pathway is associated with resistance of human breast carcinoma MCF7 cells to tumor necrosis. J Biol Chem 1997; 272: 6918–6926.

    PubMed  Google Scholar 

  28. Wissing D, Mouritzen H, Egeblad M, et al. Involvement of caspase-dependent activation of cytosolic phospholipase A2 in tumor necrosis factor-induced apoptosis. Proc Natl Acad Sci USA 1997; 94: 5073–5077.

    Google Scholar 

  29. Jaattela M, Wssing D, Kokholm K, et al. HSP70 exerts its antiapoptotic function downstream of caspase-3-like proteases. EMBO Journal 1998; 17: 6124–6134.

    PubMed  Google Scholar 

  30. Devalck D, Vercammen D, Fiers W, et al. Differential activation of phospholipases during necrosis or apoptosis—A comparative study using tumor necrosis factor and anti-fas antibodies. J Cell Biochem 1998; 71: 392–399.

    PubMed  Google Scholar 

  31. Zhang JD, Driscoll TA, Hannun YA, et al. Regulation of membrane release in apoptosis. Biochem J 1998; 334: 479–485.

    PubMed  Google Scholar 

  32. Wyllie AH. Apoptosis and carcinogenesis. Eur J Cell Biol 1997; 73: 189–197.

    PubMed  Google Scholar 

  33. Diez E, Mong S. Purification of a phospholipase A2 from human monocytic leukemic U937 cells. J Biol Chem 1990; 265: 14654–14661.

    PubMed  Google Scholar 

  34. Takayama K, Kudo I, Kim DK, et al. Purification and characterization of human platelet phospholipase A2 which preferentially hydrolyzes an arachidonoyl residue. FEBS 1991; 282: 326–330.

    Google Scholar 

  35. Kudo I, Murakami M, Hara S, et al. Mammalian non-pancreatic phospholipases A2. Biochim Biophys Acta 1993; 117: 217–231.

    Google Scholar 

  36. Vadas P, Browning J, Edelson J, et al. Extracellular phospholipase A2 expression and inflammation: The relationship with associated disease states. J Lipid Mediat 1993; 8: 1–30.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Zoology, Brigham, Young University, Provo, Utah, USA

    H. A. Wilson

  2. Department of Microbiology, Brigham Young University, Provo, Utah, USA

    D. V. Allred & K. O'Neill

  3. Department of Zoology, Brigham, Young University, Provo, Utah, USA

    J. D. Bell

Authors
  1. H. A. Wilson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. D. V. Allred
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. K. O'Neill
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J. D. Bell
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wilson, H.A., Allred, D.V., O'Neill, K. et al. Activities and interactions among phospholipases A2 during thapsigargin-induced S49 cell death. Apoptosis 5, 389–396 (2000). https://doi.org/10.1023/A:1009647912056

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1009647912056

Search

Navigation