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Caspase-3 activation is an early event and initiates apoptotic damage in a human leukemia cell line

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Abstract

Many apoptotic pathways culminate in the activation of caspase cascades usually triggered by the apical caspases-8 or -9. We describe a paradigm where apoptosis is initiated by the effector caspase-3. Diethylmaleate (DEM)-induced apoptotic damage in Jurkat cells was blocked by the anti-apoptotic protein Bcl-2, whereas, a peptide inhibitor of caspase-3 but not caspase-9 blocked DEM-induced mitochondrial damage. Isogenic Jurkat cell lines deficient for caspase-8 or the adaptor FADD (Fas associated death domain) were not protected from DEM-induced apoptosis. Caspase-3 activation preceded that of caspase-9 and initial processing of caspase-3 was regulated independent of caspase-9 and Bcl-2. However, inhibitors of caspase-9 or caspase-6 regulated caspase-3 later in the pathway. We explored the mechanism by which caspase-3 processing is regulated in this system. DEM triggered a loss of Erk-1/2 phosphorylation and XIAP (X-linked inhibitor of apoptosis protein) expression. The phorbol ester PMA activated a MEK-dependent pathway to block caspase-3 processing and cell death. Constitutively active MEK-1 (CA-MEK) upregulated XIAP expression and exogenous XIAP inhibited DEM-induced apoptotic damage. Thus, we describe a pathway where caspase-3 functions to initiate apoptotic damage and caspase-9 and caspase-6 amplify the apoptotic cascade. Further, we show that MEK may regulate caspase-3 activation via the regulation of XIAP expression in these cells.

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References

  1. Nicholson DW, Thornberry NA. Caspases: Killer proteases. Trends Biochem Sci 1997; 22: 299-306.

    Google Scholar 

  2. Slee EA, Adrain C, Martin CJ. Serial killers: Ordering caspase activation events in apoptosis. Cell Death Differ 1999; 6: 1067-1074.

    Google Scholar 

  3. Ashkenazi A, Dixit VM. Death receptors: Signaling and modulation. Science 1998; 281: 1305-1308.

    Google Scholar 

  4. Green DR, Reed JC. Mitochondria and apoptosis. Science 1998; 281: 1309-1312.

    Google Scholar 

  5. Slee EA, Harte MT, Kluck RM, et al. Ordering the cytochrome c-initiated cascade: Hierarchical activation of caspases-2,-3,-6,-7,-8 and-10 in a caspase-9 dependent manner. J Cell Biology 1999; 144: 285-291.

    Google Scholar 

  6. Van de Craen M, Declercq W, Van den Brande I, Fiers W, Vandenabeele P. The proteolytic procaspase activation network: An in vitro analysis. Cell Death and Differ 1999; 6: 1117-1124.

    Google Scholar 

  7. Komoriya A, Packard BZ, Brown MJ, Wu M-L, Henkart PA. Assessment of caspase activities in intact apoptotic thymocytes using cell-permeable flurogenic caspase substrates. J Exp Med 2000; 191: 1819-1828.

    Google Scholar 

  8. Wyllie AH. Apoptosis: An overview. Br Med Bull 1997; 53: 451-465.

    Google Scholar 

  9. Allsopp TE, McLuckie J, Kerr LE, Macleod M, Sharkey J, Kelly JS. Caspase 6 activity initiates caspase 3 activation in cerebellar granule cell apoptosis. Cell Death and Differ 2000; 7: 984-993.

    Google Scholar 

  10. Xanthoudakis S, Roy S, Rasper D, et al. Hsp60 accelerates the maturation of pro-caspase-3 by upstream activator proteases during apoptosis. EMBO J 1999; 18: 2049-2056.

    Google Scholar 

  11. Srinivasula SM, Fernandes-Alnemri T, Zangrilli J, et al. The Ced-3/interleukin 1beta converting enzyme-like homologue Mch6 and the lamin-cleaving enzyme Mch2alpha are substrates for the apoptotic mediator CPP32. J Biol Chem 1996; 271: 27099-27106.

    Google Scholar 

  12. Rao RV, Castro-Obregon S, Frankowski H, et al. Coupling endoplasmic reticulum stress to the cell death program. An Apaf-1 independent intrinsic pathway. J Biol Chem 2002; 277: 21836-21842.

    Google Scholar 

  13. Varghese J, Chattopadhyay S, Sarin A. Inhibition of p38 kinase reveals a Tumour Necrosis Factor alpha (TNF)-mediated, caspase-dependent, apoptotic death pathway in a human myelomonocyte cell line. J Immunol 2001; 166: 6570-6577.

    Google Scholar 

  14. Vander Heiden MG, Thompson CB. Bcl-2 proteins: Regulators of apoptosis or mitochondrial apoptosis? Nature Cell Biol 1999; 1: E209-E216.

    Google Scholar 

  15. Hu Y, Benedict MA, Ding L, Núñez G. Role of cytochrome c and dATP/ATP hydrolysis in Apaf-1 mediated casapse-9 activation and apoptosis. EMBO J 1999; 18: 3586-3595.

    Google Scholar 

  16. Ueda S, Nakamura H, Masutani H, et al. Redox regulation of caspase-3(-like) protease activity: Regulatory roles of thioredoxin and cytochrome c. J Immunol 1998; 161: 6689-6695.

    Google Scholar 

  17. Seol DW, Billiar TR. A caspase-9 variant missing the catalytic site is an endogenous inhibitor of apoptosis. J Biol Chem 1999; 274: 2072-2076.

    Google Scholar 

  18. Srinivasula SM, Ahmad M, Guo Y, et al. Identification of an endogenous dominant-negative short isoform of caspase-9 that can regulate apoptosis. Cancer Res 1999; 59: 999-1002.

    Google Scholar 

  19. Varghese J, Radhika G, Sarin A. The role of calpain in caspase activation during etoposide induced apoptosis in T cells. Eur J Immunol 2001; 31: 2035-2041.

    Google Scholar 

  20. Favata MF, Horiuchi KY, Manos EJ, et al. Identification of a novel inhibitor of mitogen-activated protein kinase kinase. J Biol Chem 1998; 273: 18623-18632.

    Google Scholar 

  21. Ronan N, Coffey T, William R, et al. Thiol-mediated apoptosis in prostate carcinoma cells. Cancer 2000; 88: 2092-2104.

    Google Scholar 

  22. Wolf BB, Schuler M, Echeverri F, Green DR. Caspase-3 is the primary activator of apoptotic DNA fragmentation via DNA fragmentation factor-45/inhibitor of caspase-activated DNAse inactivation. J Biol Chem 1999; 274: 30651-30656.

    Google Scholar 

  23. Fahey RJ, Doseff AI, Wewers MD. Spontaneous human monocyte apoptosis utilises a caspase-3-dependent pathway that is blocked by endotoxin and is independent of caspase-1. J Immunol 1999; 163: 1755-1762.

    Google Scholar 

  24. Kirsch DG, Doseff A, Chau BN, et al. Caspase-3-dependent cleavage of Bcl-2 promotes release of cytochrome c. J Biol Chem 1999; 274: 21155-21161.

    Google Scholar 

  25. Blane C, Deveraus QL, Krajewski S, et al. Caspase-3 is essential for procaspase-9 processing and cisplatin-induced apoptosis of MCF-7 breast cancer cells. Cancer Res 2000; 60: 4386-4390.

    Google Scholar 

  26. Tartier L, MacCarey YL, Biaglow JE, Kochevar IE, Held KD. Apoptosis induced by dithiothreitol in HL-60 cells shows early activation of caspase 3 and is independent of mtiochondria. Cell Death and Differ 2000; 7: 1002-1010.

    Google Scholar 

  27. Roy S, Bayly CI, Gareau Y, Hotzager VM, et al. Maintenance of caspase-3 proenzyme dormancy by an intrinsic “safety catch” regulatory tripeptide. Proc Natnl Acad Sci USA 2001; 98: 6132-6137.

    Google Scholar 

  28. Nakagawa T, Yuan J. Cross-talk between two cysteine protease families. Activation of caspase-12 by calpain in apoptosis. J Cell Biol. 2000; 150: 887-894.

    Google Scholar 

  29. Varghese J, Sade H, Vandenabeele P, Sarin A. Head involution (Hid) triggered apoptosis requires caspase-8 but not FADD(Fas associated death domain) and is regulated by the extracellular signal-related kianse-I/2 (Erk) in mammalian cells. J Biol Chem 2002; 277: 35097-35104.

    Google Scholar 

  30. Pardot OE, Arcaro A, Salerno G, Raguz S, Downward J, Seckl MJ. Fibroblast growth factor-2 induces translational regulation of Bcl-xL and Bcl-2 via a MEK-dependent pathway. J Biol Chem 2002; 277: 12040-12046.

    Google Scholar 

  31. Boucher MJ, Morisset J, Vachon PH, Reed JC, Laine J, Rivard N. MEK/ERK signaling pathway regulates the expression of Bcl-2, Bcl-X(L), and Mcl-1 and promotes survival of human pancreatic cancer cells. J Cell Biochem 2000; 79: 355-369.

    Google Scholar 

  32. Scheid MP, Duronio V. Dissociation of cytokine-induced phosphorylation of Bad and activation of PKB/akt: Involvement of MEK upstream of Bad phosphorylation. Proc Natnl Acad Sci USA 1998; 95: 7439-7444.

    Google Scholar 

  33. Tashker JS, Olsen M, Kornbluth S. Post-cytochrome c protection from apoptosis conferred by a MAPK pathway in Xenopus egg extracts. Mol Biol Cell 2002; 13: 393-401.

    Google Scholar 

  34. Erhardt P, Schremser EJ, Cooper GM. B-Raf inhibits programmed cell death downstream of cytochrome c release from mitochondria by activating the MEK/Erk pathway. Mol Cell Biol 1999; 19: 5308-5315.

    Google Scholar 

  35. Deveraux QL, Reed JC. IAP family proteins-suppressors of apoptosis. Genes Dev 1999; 13: 239-252.

    Google Scholar 

  36. Stennicke HR, Ryan CA, Salvesen GS. Reprieval from execution: The molecular basis of caspase inhibition. Trends Biochem Sci 2002; 27: 94-101.

    Google Scholar 

  37. Pae HO, Yoo JC, Choi BM, Lee EJ, Song YS, Chung HT. 12-Otetradecanoyl phorbol 13-acetate, protein kinase C (PKC) activator, protects human leukemia HL-60 cels from taxol-induced apoptosis: Possible role for extracellular signal-regulated kinase. Immunopharmacol Immunotoxicol 2000; 22: 61-73.

    Google Scholar 

  38. Zhuang S, Lynch MC, Kochevar IE. Activation of protein kinase C is required for protection of cells against apoptosis induced by singlet oxygen. FEBS Lett 1998; 437: 158-162.

    Google Scholar 

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Authors and Affiliations

  1. National Centre for Biological Sciences, New Bellary Road, UAS-GKVK Campus, Bangalore, 560065, Karnataka, India

    J. Varghese, N. S. Khandre & A. Sarin

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  1. J. Varghese
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  2. N. S. Khandre
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  3. A. Sarin
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Varghese, J., Khandre, N.S. & Sarin, A. Caspase-3 activation is an early event and initiates apoptotic damage in a human leukemia cell line. Apoptosis 8, 363–370 (2003). https://doi.org/10.1023/A:1024121017841

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