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Caspase-3/CPP32-like activity is not sufficient to mediate apoptosis in an IL-2 dependent T cell line

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Abstract

CTLL cells undergo apoptosis when cultured in the absence of IL-2. The IL-1β-converting-enzyme (ICE)/ caspase family has been implicated as an integral component of some forms of apoptosis. Numerous members of the caspase family have been identified, and it appears as if caspase-3/CPP32 plays a critical role. Previously we demonstrated that ICE/caspase-1 expression increases in CTLL cells during apoptosis; however, inhibition of ICE activity did not abrogate apoptotic death. The purpose of this report is to determine if other members of the caspase family are involved in T cell apoptosis induced by growth factor starvation. We show that cytosolic CPP32-like activity, as measured by the cleavage of DEVD-pNA and poly(ADP-ribose) polymerase (PARP), increases during apoptosis following growth factor deprivation. Cytosolic CPP32-like activity is inhibited in cells treated with the broad spectrum ICE family inhibitor boc-aspartyl(OMe)-fluoromethylketone (D-FMK) and by VAD-FMK and DEVD-FMK which have greater specificity for CPP32-like ICE homologs; however, only the broad spectrum ICE inhibitor D-FMK inhibited apoptosis. Our results suggest that apoptosis induced by growth factor deprivation involves the caspase family, but increased CPP32-like activity is not sufficient to mediate apoptosis induced by IL-2 starvation.

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References

  1. Cohen J. Programmed cell death in the immune system. Adv Immunol 1991; 50: 55-85.

    Google Scholar 

  2. Lenardo M. Interleukin-2 programs mouse alpha beta T lymphocytes for apoptosis. Nature 1991; 353: 858-861.

    Google Scholar 

  3. Ameisen J, Capron A. Cell dysfunction and depletion in AIDS: the programmed cell death hypothesis. Immunol Today 1991; 12: 102-105.

    Google Scholar 

  4. Cheng J, Zhou T, Liu C, et al. A soluble form of the Fas molecule protects cells from Fas-mediated apoptosis. Science 1994; 263: 1759-1762.

    Google Scholar 

  5. Meyaard L, Otto S, Jonker R, Mijnster M, Keet R, Miedema F. Programmed death of T cells in HIV-1 infection. Science 1992; 257: 217-219.

    Google Scholar 

  6. Biedler D, Tewari M, Friesen P, Poirier G, Dixit V. The baculovirus p35 protein inhibits Fas-and tumor necrosis factor-induced apoptosis. J Biol Chem 1995; 270: 16526-16528.

    Google Scholar 

  7. Cain K, Inayat-Hussain S, Couet C, Cohen G. A cleavage-site-directed inhibitor of interleukin-1β-converting enzyme-like proteases inhibits apoptosis in primary cultures of rat hepatocytes. Biochem J 1996; 314: 27-32.

    Google Scholar 

  8. Gagliardini V, Fernandez P-A, Lee R, et al. Prevention of vertebrate neuronal death by the crmA gene. Science 1994; 263: 826-828.

    Google Scholar 

  9. Fearnhead H, Dinsdale D, Cohen G. An IL-1β-converting enzyme-like protease is a common mediator of apoptosis in thymocytes. FEBS Lett 1995; 375: 283-288.

    Google Scholar 

  10. Kumar S, Kinoshita M, Noda M, Copeland N, Jenkins N. Induction of apoptosis by the mouse Nedd2 gene, which encodes a protein similar to the product of the Caenorhabditis elegans cell death gene ced-3 and the mammalian IL-1β-converting enzyme. Genes Dev 1994; 8: 1613-1626.

    Google Scholar 

  11. Milligan C, Prevette D, Yaginuma H, et al. Peptide inhibitors of the ICE protease family arrest programmed cell death of motorneurons in vivo and in vitro. Neuron 1995; 15: 385-393.

    Google Scholar 

  12. Enari M, Hug H, Nagata S. Involvement of an ICE-like protease in Fas-mediated apoptosis. Nature 1995; 375: 78-81.

    Google Scholar 

  13. Zhu H, Fearnhead H, Cohen G. An ICE-like protease is a common mediator of apoptosis induced by diverse stimuli in human monocytic THP.1 cells. FEBS Lett 1995; 374: 303-308.

    Google Scholar 

  14. Yuan J, Shaham S, Ledoux S, Ellis H, Horvitz H. The C. elegans cell death gene ced-3 encodes a protein similar to mammalian interleukin-1β-converting enzyme. Cell 1993; 75: 641-652.

    Google Scholar 

  15. Miura M, Friedlander R, Yuan J. Tumor necrosis factor-induced apoptosis is mediated by a CrmA-sensitive cell death pathway. Proc Natl Acad Sci USA 1995; 92: 8318-8322.

    Google Scholar 

  16. Schlegel J, Peters I, Orrenius S. Isolation and partial characterization of a protease involved in Fas-induced apoptosis. FEBS Lett 1995; 364: 139-142.

    Google Scholar 

  17. Slee E, Zhu H, Chow S, MacFarlane M, Nicholson D, Cohen G. Benzyloxycarbonyl-Val-Ala-Asp (OMe) fluoromethylketone (Z-VAD.FMK) inhibits apoptosis by blocking the processing of CPP32. Biochem J 1996; 315: 21-24.

    Google Scholar 

  18. Xue D, Shaham S, Horvitz H. The Caenorhabditis elegans cell-death protein CED-3 is a cysteine protease with substrate specificities similar to those of the human CPP32 protease. Genes Dev 1996; 10: 1073-1083.

    Google Scholar 

  19. Jacobson M, Weil M, Raf M. Role of Ced-3/ICE-family proteases in staurosporine-induced programmed cell death. J Cell Biol 1996; 133: 1041-1051.

    Google Scholar 

  20. Wang L, Miura M, Bergeron L, Zhu H, Yuan J. Ich-1, an Ice/ced-3-related gene, encodes both positive and negative regulators of programmed cell death. Cell 1994; 78: 739-750.

    Google Scholar 

  21. Wang S, Miura M, Jung Y-K, et al. Identification and characterization of Ich-3, a member of the interleukin-1β-converting enzyme (ICE)/Ced-3 family and an upstream regulator of ICE. J Biol Chem 1996; 271: 20580-20587.

    Google Scholar 

  22. Mashima T, Naito M, Kataoka S, Kawai H, Tsuruo T. Aspartate-based inhibitor of interleukin-1β-converting enzyme prevents antitumor agent-induced apoptosis in human myeloid leukemia U937 cells. Biochem Biophys Res Com 1995; 209: 907-915.

    Google Scholar 

  23. Hasegawa J, Kamada S, Kamiike W, et al. Involvement of CPP32/Yama(-like) proteases in Fas-mediated apoptosis. Cancer Res 1996; 56: 1713-1718.

    Google Scholar 

  24. Nicholson D, Ali A, Thornberry N, et al. Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis. Nature 1995; 376: 37-43.

    Google Scholar 

  25. Duan H, Chinnaiyan A, Hudson P, Wing J, He W, Dixit V. ICE-LAP3, a novel mammalian homologue of the Caenorhabditis elegans cell death protein Ced-3 is activated during Fas-and tumor necrosis factor-induced apoptosis. J Biol Chem 1996; 271: 1621-1625.

    Google Scholar 

  26. Duan H, Orth K, Chinnaiyan A, et al. ICE-Lap6, a novel member of the ICE/Ced-3 gene family, is activated by the cytotoxic T cell protease granzyme B. J Biol Chem 1996; 271: 16720-16724.

    Google Scholar 

  27. Faucheu C, Diu A, Chan A, et al. A novel human protease similar to the interleukin-1β converting enzyme induces apoptosis in transfected cells. EMBO J 1995; 14: 1914-1922.

    Google Scholar 

  28. Faucheu C, Blanchet AM, Collard-Dutilleul V, Lalanne JL, Diu-Hercend A. Identification of a cysteine protease closely related to interleukin-1β-converting enzyme. Eur J Biochem 1996; 236: 207-213.

    Google Scholar 

  29. Fernandes-Alnemri T, Litwack, G, Alnemri E. Mch2, a new member of the apoptotic Ced-3/Ice cysteine protease gene family. Cancer Res 1995; 55: 2737-2742.

    Google Scholar 

  30. Fernandes-Alnemri T, Takahashi A, Armstrong R, et al. Mch3, a novel human apoptotic cysteine protease related to CPP32. Cancer Res 1996; 55: 6045-6052.

    Google Scholar 

  31. Kamens J, Paskind M, Hugunin M, et al. Identification and characterization of ICH-2, a novel member of the interleukin-1β-converting enzyme family of cysteine proteases. J Biol Chem 1995; 270: 15250-15256.

    Google Scholar 

  32. Lippke J, Gu Y, Sarnecki C, Caron P, M Su. Identification and characterization of CPP32/Mch2 homolog 1, a novel cysteine protease similar to CPP32. J Biol Chem 1996; 271: 1825-1828.

    Google Scholar 

  33. Pai J, Brown M, Goldstein J. Purification and cDNA cloning of a second apoptosis-related cysteine protease that cleaves and activates sterol regulatory element binding proteins. Proc Natl Acad Sci USA 1996; 93: 5437-5442.

    Google Scholar 

  34. Wang X, Pai J, Wiedenfeld E, et al. Purification of an interleukin-1β converting enzyme-related cysteine protease that cleaves sterol regulatory element-binding proteins between the leucine zipper and transmembrane domains. J Biol Chem 1995; 270: 18044-18050.

    Google Scholar 

  35. Na S, Chuang TS, Cunningham A, et al. D4-GDI, a substrate of CPP32, is proteolyzed during Fas-induced apoptosis. J Biol Chem 1996; 271: 11209-11213.

    Google Scholar 

  36. Nath R, Raser K, Stafford D, et al. Non-erythroid α-spectrin breakdown by calpain and interleukin 1β-converting-enzyme-like protease(s) in apoptotic cells: contributory roles of both protease families in neuronal apoptosis. Biochem J 1996; 319: 683-690.

    Google Scholar 

  37. Tewari M, Quan L, O'Rourke K, et al. Yama/CPP32β, a mammalian homolog of CED-3, is a crmA-inhibitable protease that cleaves the death substrate poly(ADP-ribose) polymerase. Cell 1995; 81: 801-809.

    Google Scholar 

  38. Casiola-Rosen L, Nicholson D, Chong T, et al. Apopain/CPP32 cleaves proteins that are essential for cellular repair: A fundamental principle of apoptotic death. J Exp Med 1996; 183: 1957-1964.

    Google Scholar 

  39. Darmon A, Nicholson D, Bleackley R. Activation of the apoptotic protease CPP32 by cytotoxic T-cell-derived granzyme B. Nature 1995; 377: 446-448.

    Google Scholar 

  40. Duke R, Cohen J. IL-2 addiction: withdrawal of growth factor activates a suicide program in dependent T cells. Lymphokine Res 1986; 5: 289-299.

    Google Scholar 

  41. Deng G, Podack E. Suppression of apoptosis in a cytotoxic T-cell line by interleukin 2-mediated gene transcription and deregulated expression of the proto-oncogene bcl-2. Proc Natl Acad Sci USA 1993; 90: 2189-2193.

    Google Scholar 

  42. Vasilakos J, Ghayur T, Carroll R, et al. IL-1β converting enzyme (ICE) is not required for apoptosis induced by lymphokine deprivation in an IL-2-dependent T cell line. J Immunol 1995; 155: 3433-3442.

    Google Scholar 

  43. Schlegel J, Peters I, Orrenius S, et al. CPP32/apopain is a key interleukin-1β converting enzyme-like protease involved in Fas-mediated apoptosis. J Biol Chem 1996; 271: 1841-1844.

    Google Scholar 

  44. Bump N, Hackett M, Hugunin M, et al. Inhibition of ICE family proteases by baculovirus antiapoptotic protein p35. Science 1995; 269: 1885-1888.

    Google Scholar 

  45. Schmaier A, Bradford H, Lundberg D, Farber A, Colman R. Membrane expression of platelet calpain. Blood 1990; 75: 1273-1281.

    Google Scholar 

  46. Chu-Ping M, Slaughter C, DeMartino G. Purification and Characterization of a protein inhibitor of the 20S proteasome (macropain). Biochem Biophys Acta 1992; 1119: 303-311.

    Google Scholar 

  47. Schulze-Osthoff K, Krammer P, Droge W. Divergent signaling via APO-1/Fas and the TNF receptor, two homologous molecules involved in physiological cell death. EMBO J 1994; 13: 4587-4596.

    Google Scholar 

  48. Kuida K, Lippke J, Ku G, et al. Altered cytokine export and apoptosis in mice deficient in IL-1β converting enzyme. Science 1995; 267: 2000-2003.

    Google Scholar 

  49. Thornberry N, Bull H, Calaycay J, et al. A novel heterodimeric cysteine protease is required for interleukin-1β processing in monocytes. Nature 1992; 356: 768-774.

    Google Scholar 

  50. Fernandes-Alnemri T, Litwack G, Alnemri E. CPP32, a novel human apoptotic protein with homology to Caenorhabditis elegans cell death protein Ced-3 and mammalian interleukin-1β-converting enzyme. J Biol Chem 1994; 269: 30761-30764.

    Google Scholar 

  51. Kaufmann S, Desnoyers S, Ottaviano Y, Davidson N, Poirier G. Specific proteolytic cleavage of poly(ADP-ribose) polymerase: an early marker of chemotherapy-induced apoptosis. Cancer Res 1993; 53: 3976-3985.

    Google Scholar 

  52. Lazebnik Y, Kaufmann S, Desnoyers S, Poirier G, Earnshaw W. Cleavage of poly(ADP-ribose) polymerase by a proteinase with properties like ICE. Nature 1994; 371: 346-347.

    Google Scholar 

  53. Armstrong R, Aja T, Xiang J, et al. Fas-induced activation of the cell death-related protease CPP32 is inhibited by Bcl-2 and by ICE family protease inhibitors. J Biol Chem 1996; 271: 16850-16855.

    Google Scholar 

  54. Deshmukh M, Vasilakos J, Deckwerth T, Lampe P, Shivers B, Johnson E. Genetic and metabolic status of NGF-deprived sympathetic neurons saved by an inhibitor of ICE-family proteases. J Cell Biol 1996; 135: 1341-1354.

    Google Scholar 

  55. Boudreau N, Sympson C, Werb Z, Bissell M. Suppression of ICE and apoptosis in mammary epithelial cells by extracellular matrix. Science 1995; 267: 891-893.

    Google Scholar 

  56. Wilson K, Black J, Thompson J, et al. Structure and mechanism of interleukin-1β converting enzyme. Nature 1994; 370: 270-275.

    Google Scholar 

  57. Walker N, Talanian R, Brady K, et al. Crystal structure of the cysteine protease interleukin-1β-converting enzyme: a (p20/p10)2 homodimer. Cell 1994; 78: 343-352.

    Google Scholar 

  58. Rauber P, Angliker H, Walker B, Shaw E. The synthesis of peptidylfluoromethanes and their properties as inhibitors of serine proteinases and cysteine proteinases. Biochem J 1986; 239: 633-640.

    Google Scholar 

  59. Squier M, Miller A, Malkinson A, Cohen J. Calpain activation in apoptosis. J Cell Physiol 1994; 159: 229-237.

    Google Scholar 

  60. Enari M, Talanian R, Wong W, Nagata S. Sequential activation of ICE-like and CPP32-like proteases during Fas-mediated apoptosis. Nature 1996; 380: 723-726.

    Google Scholar 

  61. Miura M, Zhu H, Rotello R, Hartwieg E, Yuan J. Induction of apoptosis in fibroblasts by IL-1β converting enzyme, a mammalian homolog of the C. elegans cell death gene ced-3. Cell 1993; 75: 653-660.

    Google Scholar 

  62. Dolle R, Hoyer D, Prasad C, et al. P1 aspartate-based peptide alpha-((2,6-dichlorobenzoyl)oxy)methyl ketones as potent time-dependent inhibitors of interleukin-1β-converting enzyme. J Med Chem 1994; 37: 563-564.

    Google Scholar 

  63. Sarin A, Wu ML, Henkart P. Different interleukin-1 beta converting enzyme (ICE) family protease requirements for the apoptotic death of T lymphocytes triggered by diverse stimuli. J Exp Med 1996; 184: 2445-2450.

    Google Scholar 

  64. Martin S, Amarante-Mendes G, Shi L, et al. The cytotoxic cell protease granzyme B initiates apoptosis in a cell-free system by proteolytic processing and activation of the ICE/CED-3 family protease, CPP32, via a novel two-step mechanism. EMBO J 1996; 15: 2407-2416.

    Google Scholar 

  65. An S, Knox K. Ligation of CD40 rescues Ramos-Burkitt lymphoma B cells from calcium ionophore-and antigen receptor-triggered apoptosis by inhibiting activation of the cysteine protease CPP32/Yama and cleavage of its substrate PARP. FEBS Lett 1996; 386: 115-122.

    Google Scholar 

  66. Chow S, Weis M, Kaas G, Homstrom T, Eriksson J, Orrenius S. Involvement of multiple proteases during Fas-mediated apoptosis in T lymphocytes. FEBS Lett 1995; 364: 134-138.

    Google Scholar 

  67. Troy C, Stefanis L, Prochiantz A, Greene L, Shelanski M. The contrasting roles of ICE family proteases and interleukin-1β in apoptosis induced by trophic factor withdrawal and by copper/zinc superoxide dismutase down-regulation. Proc Natl Acad Sci USA 1996; 93: 5635-5640.

    Google Scholar 

  68. Kumar S. Inhibition of apoptosis by the expression of antisense Nedd2. FEBS Lett 1995; 368: 69-72.

    Google Scholar 

  69. Kuida K, Zheng T, Na S, et al. Decreased apoptosis in the brain and premature lethality in CPP32-deficient mice. Nature 1996; 384: 368-372.

    Google Scholar 

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Authors and Affiliations

  1. Department of Cell Biology, Parke-Davis Pharmaceutical Research Company, Division of Warner-Lambert, Ann Arbor, USA

    J. P. Vasilakos, T. Lynch & B. D. Shivers

  2. Department of Neurology, University of Michigan, Ann Arbor, USA

    T. Lynch, D. A. Giegel & M. Santoro

  3. BASF Bioresearch Corporation, Worcester, USA

    T. Ghayur

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  1. J. P. Vasilakos
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  2. T. Lynch
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  4. D. A. Giegel
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Vasilakos, J.P., Lynch, T., Ghayur, T. et al. Caspase-3/CPP32-like activity is not sufficient to mediate apoptosis in an IL-2 dependent T cell line. Apoptosis 2, 289–303 (1997). https://doi.org/10.1023/A:1026489104188

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