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PMID: 7489708 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Cell-free reconstitution of Fas-, UV radiation- and ceramide-induced apoptosis.

The EMBO journal ·Vol. 14 ·No. 21 ·1995-11-01 ·Pages 5191-200

Martin SJ, Newmeyer DD, Mathias S, Farschon DM, Wang HG, Reed JC, Kolesnick RN, Green DR

Abstract

Cell-free systems are valuable tools for the dissection of complex cellular processes. Here we show that cytoplasmic extracts from cells exposed to anti-Fas antibody or UV radiation contain an activity capable of reproducing morphological changes typical of apoptosis in nuclei added to these extracts, as well as internucleosomal cleavage of DNA and proteolysis of a protein known to be cleaved during the apoptosis of intact cells. Extracts from control cell populations were inactive in this respect. These effects were partly blocked by the addition of purified Bcl-2 protein or a competitive inhibitor peptide of interleukin-1 beta-converting enzyme to the extracts. Furthermore, apoptotic activity was induced in cytoplasmic extracts from untreated cells by the addition of ceramide, a lipid second messenger implicated recently in apoptosis signaling. These extracts should prove highly useful in the dissection of molecular events that occur during apoptosis.

MeSH Terms
Animals Antibodies/immunology Apoptosis/drug effects,physiology,radiation effects Caspase 1 Cell Nucleus/drug effects,physiology,radiation effects Cell-Free System Ceramides/pharmacology Cysteine Endopeptidases/physiology DNA/analysis Humans Proto-Oncogene Proteins/physiology Proto-Oncogene Proteins c-bcl-2 Ultraviolet Rays fas Receptor/immunology
Chemicals
Antibodies Ceramides Proto-Oncogene Proteins Proto-Oncogene Proteins c-bcl-2 fas Receptor DNA Cysteine Endopeptidases Caspase 1
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Martin S J
Division of Cellular Immunology, La Jolla Institute for Allergy and Immunology, CA 92037, USA.
Newmeyer D D
Mathias S
Farschon D M
Wang H G
Reed J C
Kolesnick R N
Green D R
References (72)
72 references, click to expand
  1. The C. elegans cell death gene ced-3 encodes a protein similar to mammalian interleukin-1 beta-converting enzyme.
    Cell. 1993 Nov 19;75(4):641-52 PMID: 8242740
  2. Gene induction by gamma-irradiation leads to DNA fragmentation in lymphocytes.
    J Immunol. 1987 Nov 15;139(10):3199-206 PMID: 3680944
  3. Ceramide synthase mediates daunorubicin-induced apoptosis: an alternative mechanism for generating death signals.
    Cell. 1995 Aug 11;82(3):405-14 PMID: 7634330
  4. Cell nucleus and DNA fragmentation are not required for apoptosis.
    J Cell Biol. 1994 Oct;127(1):15-20 PMID: 7523418
  5. Tumor necrosis factor can induce both apoptic and necrotic forms of cell lysis.
    J Immunol. 1988 Oct 15;141(8):2629-34 PMID: 3171180
  6. Protease activation during apoptosis: death by a thousand cuts?
    Cell. 1995 Aug 11;82(3):349-52 PMID: 7634323
  7. Social controls on cell survival and cell death.
    Nature. 1992 Apr 2;356(6368):397-400 PMID: 1557121
  8. Induction of apoptotic DNA damage and cell death by activation of the sphingomyelin pathway.
    Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):73-7 PMID: 8278410
  9. Ionizing radiation acts on cellular membranes to generate ceramide and initiate apoptosis.
    J Exp Med. 1994 Aug 1;180(2):525-35 PMID: 8046331
  10. Prevention of vertebrate neuronal death by the crmA gene.
    Science. 1994 Feb 11;263(5148):826-8 PMID: 8303301
  11. Thymocyte apoptosis induced by p53-dependent and independent pathways.
    Nature. 1993 Apr 29;362(6423):849-52 PMID: 8479523
  12. Induction of apoptosis in fibroblasts by IL-1 beta-converting enzyme, a mammalian homolog of the C. elegans cell death gene ced-3.
    Cell. 1993 Nov 19;75(4):653-60 PMID: 8242741
  13. Cell-free apoptosis in Xenopus egg extracts: inhibition by Bcl-2 and requirement for an organelle fraction enriched in mitochondria.
    Cell. 1994 Oct 21;79(2):353-64 PMID: 7954801
  14. FAS-induced apoptosis is mediated via a ceramide-initiated RAS signaling pathway.
    Immunity. 1995 Apr;2(4):341-51 PMID: 7536620
  15. Egg extracts for nuclear import and nuclear assembly reactions.
    Methods Cell Biol. 1991;36:607-34 PMID: 1811153
  16. Dicing with death: dissecting the components of the apoptosis machinery.
    Trends Biochem Sci. 1994 Jan;19(1):26-30 PMID: 8140617
  17. HIV-1 infection of human CD4+ T cells in vitro. Differential induction of apoptosis in these cells.
    J Immunol. 1994 Jan 1;152(1):330-42 PMID: 7902858
  18. Suppression of ICE and apoptosis in mammary epithelial cells by extracellular matrix.
    Science. 1995 Feb 10;267(5199):891-3 PMID: 7531366
  19. Apoptosis reduces both the in vitro replication and the in vivo infectivity of a baculovirus.
    J Virol. 1993 Jul;67(7):3730-8 PMID: 8510202
  20. Bcl-2 oncoprotein blocks chemotherapy-induced apoptosis in a human leukemia cell line.
    Blood. 1993 Jan 1;81(1):151-7 PMID: 8417786
  21. Purification of three cytotoxic lymphocyte granule serine proteases that induce apoptosis through distinct substrate and target cell interactions.
    J Exp Med. 1992 Dec 1;176(6):1521-9 PMID: 1460416
  22. Ceramide stimulates a cytosolic protein phosphatase.
    J Biol Chem. 1992 Mar 15;267(8):5048-51 PMID: 1312082
  23. Bcl-2: a repressor of lymphocyte death.
    Immunol Today. 1992 Aug;13(8):285-8 PMID: 1510811
  24. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation.
    Nature. 1980 Apr 10;284(5756):555-6 PMID: 6245367
  25. Induction of apoptosis in fibroblasts by c-myc protein.
    Cell. 1992 Apr 3;69(1):119-28 PMID: 1555236
  26. Activation of the Abelson tyrosine kinase activity is associated with suppression of apoptosis in hemopoietic cells.
    Cancer Res. 1993 Apr 15;53(8):1735-8 PMID: 8467488
  27. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics.
    Br J Cancer. 1972 Aug;26(4):239-57 PMID: 4561027
  28. Programmed cell death induced by ceramide.
    Science. 1993 Mar 19;259(5102):1769-71 PMID: 8456305
  29. Role for c-myc in activation-induced apoptotic cell death in T cell hybridomas.
    Science. 1992 Jul 10;257(5067):212-4 PMID: 1378649
  30. p53 is required for radiation-induced apoptosis in mouse thymocytes.
    Nature. 1993 Apr 29;362(6423):847-9 PMID: 8479522
  31. Mice deficient in IL-1 beta-converting enzyme are defective in production of mature IL-1 beta and resistant to endotoxic shock.
    Cell. 1995 Feb 10;80(3):401-11 PMID: 7859282
  32. Fodrin is the general spectrin-like protein found in most cells whereas spectrin and the TW protein have a restricted distribution.
    Cell. 1983 Sep;34(2):503-12 PMID: 6352052
  33. BCR-ABL maintains resistance of chronic myelogenous leukemia cells to apoptotic cell death.
    Blood. 1994 Mar 1;83(5):1179-87 PMID: 8118022
  34. Monoclonal antibody-mediated tumor regression by induction of apoptosis.
    Science. 1989 Jul 21;245(4915):301-5 PMID: 2787530
  35. Dose-dependent induction of apoptosis in human tumour cell lines by widely diverging stimuli.
    Cell Prolif. 1991 Mar;24(2):203-14 PMID: 2009322
  36. Protease inhibitors selectively block T cell receptor-triggered programmed cell death in a murine T cell hybridoma and activated peripheral T cells.
    J Exp Med. 1993 Nov 1;178(5):1693-700 PMID: 8228816
  37. Substrate recognition by ceramide-activated protein kinase. Evidence that kinase activity is proline-directed.
    J Biol Chem. 1993 Sep 25;268(27):20002-6 PMID: 8376361
  38. Bcl-2 and the regulation of programmed cell death.
    J Cell Biol. 1994 Jan;124(1-2):1-6 PMID: 8294493
  39. Ultraviolet B irradiation of human leukaemia HL-60 cells in vitro induces apoptosis.
    Int J Radiat Biol. 1991 Apr;59(4):1001-16 PMID: 1674267
  40. Ceramide: a signal for apoptosis or mitogenesis?
    J Exp Med. 1995 Jun 1;181(6):1949-52 PMID: 7759991
  41. Molecular regulation of apoptosis: genetic controls on cell death.
    Cell. 1993 Sep 10;74(5):777-9 PMID: 8104100
  42. Calpain activation in apoptosis.
    J Cell Physiol. 1994 May;159(2):229-37 PMID: 8163563
  43. Activation of Epstein-Barr virus latent genes protects human B cells from death by apoptosis.
    Nature. 1991 Feb 14;349(6310):612-4 PMID: 1705663
  44. Apoptosis and cancer: the failure of controls on cell death and cell survival.
    Crit Rev Oncol Hematol. 1995 Feb;18(2):137-53 PMID: 7695828
  45. Specific proteolytic cleavage of poly(ADP-ribose) polymerase: an early marker of chemotherapy-induced apoptosis.
    Cancer Res. 1993 Sep 1;53(17):3976-85 PMID: 8358726
  46. Activation of intracellular proteases is an early event in TNF-induced apoptosis.
    J Immunol. 1995 Feb 15;154(4):1707-16 PMID: 7836755
  47. Nuclear events of apoptosis in vitro in cell-free mitotic extracts: a model system for analysis of the active phase of apoptosis.
    J Cell Biol. 1993 Oct;123(1):7-22 PMID: 8408207
  48. The sphingomyelin pathway in tumor necrosis factor and interleukin-1 signaling.
    Cell. 1994 May 6;77(3):325-8 PMID: 8181053
  49. Fas- and tumor necrosis factor-induced apoptosis is inhibited by the poxvirus crmA gene product.
    J Biol Chem. 1995 Feb 17;270(7):3255-60 PMID: 7531702
  50. Apoptotic signaling through CD95 (Fas/Apo-1) activates an acidic sphingomyelinase.
    J Exp Med. 1994 Oct 1;180(4):1547-52 PMID: 7523573
  51. Ceramide: an intracellular signal for apoptosis.
    Trends Biochem Sci. 1995 Feb;20(2):73-7 PMID: 7701566
  52. Involvement of spectrin in the maintenance of phase-state asymmetry in the erythrocyte membrane.
    Cell. 1982 Oct;30(3):725-33 PMID: 7139713
  53. Apoptosis and programmed cell death in immunity.
    Annu Rev Immunol. 1992;10:267-93 PMID: 1590988
  54. Direct stimulation of Vav guanine nucleotide exchange activity for Ras by phorbol esters and diglycerides.
    Mol Cell Biol. 1994 Jul;14(7):4749-58 PMID: 7516472
  55. Apoptosis. The role of the endonuclease.
    Am J Pathol. 1990 Mar;136(3):593-608 PMID: 2156431
  56. Cleavage of poly(ADP-ribose) polymerase by a proteinase with properties like ICE.
    Nature. 1994 Sep 22;371(6495):346-7 PMID: 8090205
  57. Inhibition of apoptosis by BCR-ABL in chronic myeloid leukemia.
    Blood. 1994 Apr 15;83(8):2038-44 PMID: 8161775
  58. Endogenous endonuclease-induced DNA fragmentation: an early event in cell-mediated cytolysis.
    Proc Natl Acad Sci U S A. 1983 Oct;80(20):6361-5 PMID: 6312454
  59. Ich-1, an Ice/ced-3-related gene, encodes both positive and negative regulators of programmed cell death.
    Cell. 1994 Sep 9;78(5):739-50 PMID: 8087842
  60. Bcl-2 is an inner mitochondrial membrane protein that blocks programmed cell death.
    Nature. 1990 Nov 22;348(6299):334-6 PMID: 2250705
  61. Key morphological features of apoptosis may occur in the absence of internucleosomal DNA fragmentation.
    Biochem J. 1992 Sep 1;286 ( Pt 2):331-4 PMID: 1530564
  62. Specific cleavage of the 70-kDa protein component of the U1 small nuclear ribonucleoprotein is a characteristic biochemical feature of apoptotic cell death.
    J Biol Chem. 1994 Dec 9;269(49):30757-60 PMID: 7983001
  63. Programmed cell death and Bcl-2 protection in the absence of a nucleus.
    EMBO J. 1994 Apr 15;13(8):1899-910 PMID: 8168488
  64. Apoptotic death in epithelial cells: cleavage of DNA to 300 and/or 50 kb fragments prior to or in the absence of internucleosomal fragmentation.
    EMBO J. 1993 Sep;12(9):3679-84 PMID: 8253089
  65. Wild-type p53 induces apoptosis of myeloid leukaemic cells that is inhibited by interleukin-6.
    Nature. 1991 Jul 25;352(6333):345-7 PMID: 1852210
  66. Altered cytokine export and apoptosis in mice deficient in interleukin-1 beta converting enzyme.
    Science. 1995 Mar 31;267(5206):2000-3 PMID: 7535475
  67. Rodent fibroblast tumours expressing human myc and ras genes: growth, metastasis and endogenous oncogene expression.
    Br J Cancer. 1987 Sep;56(3):251-9 PMID: 3663473
  68. CPP32, a novel human apoptotic protein with homology to Caenorhabditis elegans cell death protein Ced-3 and mammalian interleukin-1 beta-converting enzyme.
    J Biol Chem. 1994 Dec 9;269(49):30761-4 PMID: 7983002
  69. A cell-killing monoclonal antibody (anti-Fas) to a cell surface antigen co-downregulated with the receptor of tumor necrosis factor.
    J Exp Med. 1989 May 1;169(5):1747-56 PMID: 2469768
  70. Proteolysis of fodrin (non-erythroid spectrin) during apoptosis.
    J Biol Chem. 1995 Mar 24;270(12):6425-8 PMID: 7534762
  71. The apoptosis endonucleases: cleaning up after cell death?
    Trends Cell Biol. 1994 Feb;4(2):37-41 PMID: 14731864
  72. Induction of apoptosis (programmed cell death) in human leukemic HL-60 cells by inhibition of RNA or protein synthesis.
    J Immunol. 1990 Sep 15;145(6):1859-67 PMID: 2167911
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1995-11-01
Pages
5191-200
Language
English
Region
England
NLM ID
8208664
PMCID
PMC394627
Subset
IM
Grants
NIGMS NIH HHS · GM52735 · United States
External Links
PubMed source PMC Full Text View journal details
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