Essential contribution of caspase 3/CPP32 to apoptosis and its associated nuclear changes.

Caspases are fundamental components of the mammalian apoptotic machinery, but the precise contribution of individual caspases is controversial. CPP32 (caspase 3) is a prototypical caspase that becomes activated during apoptosis. In this study, we took a comprehensive approach to examining the role of CPP32 in apoptosis using mice, embryonic stem (ES) cells, and mouse embryonic fibroblasts (MEFs) deficient for CPP32. CPP32(ex3-/-) mice have reduced viability and, consistent with an earlier report, display defective neuronal apoptosis and neurological defects. Inactivation of CPP32 dramatically reduces apoptosis in diverse settings, including activation-induced cell death (AICD) of peripheral T cells, as well as chemotherapy-induced apoptosis of oncogenically transformed CPP32(-/-) MEFs. As well, the requirement for CPP32 can be remarkably stimulus-dependent: In ES cells, CPP32 is necessary for efficient apoptosis following UV- but not gamma-irradiation. Conversely, the same stimulus can show a tissue-specific dependence on CPP32: Hence, TNFalpha treatment induces normal levels of apoptosis in CPP32 deficient thymocytes, but defective apoptosis in oncogenically transformed MEFs. Finally, in some settings, CPP32 is required for certain apoptotic events but not others: Select CPP32(ex3-/-) cell types undergoing cell death are incapable of chromatin condensation and DNA degradation, but display other hallmarks of apoptosis. Together, these results indicate that CPP32 is an essential component in apoptotic events that is remarkably system- and stimulus-dependent. Consequently, drugs that inhibit CPP32 may preferentially disrupt specific forms of cell death.

[1]  J. Rothman,et al.  Baculovirus p35 prevents developmentally programmed cell death and rescues a ced‐9 mutant in the nematode Caenorhabditis elegans. , 1994, The EMBO journal.

[2]  M. Salmon,et al.  Cellular environments and apoptosis: tissue microenvironments control activated T-cell death. , 1997, Immunology today.

[3]  D. Housman,et al.  p53-dependent apoptosis modulates the cytotoxicity of anticancer agents , 1993, Cell.

[4]  S. Korsmeyer,et al.  Fas-induced Activation of the Cell Death-related Protease CPP32 Is Inhibited by Bcl-2 and by ICE Family Protease Inhibitors* , 1996, The Journal of Biological Chemistry.

[5]  A. Chinnaiyan,et al.  ICE-LAP6, a Novel Member of the ICE/Ced-3 Gene Family, Is Activated by the Cytotoxic T Cell Protease Granzyme B* , 1996, The Journal of Biological Chemistry.

[6]  Scott W. Lowe,et al.  p53 is required for radiation-induced apoptosis in mouse thymocytes , 1993, Nature.

[7]  S. Nagata,et al.  Apoptosis by Death Factor , 1997, Cell.

[8]  D. Nicholson,et al.  Activation of the apoptotic protease CPP32 by cytotoxic T-cell-derived granzyme B , 1995, Nature.

[9]  H. Horvitz,et al.  C. elegans cell survival gene ced-9 encodes a functional homolog of the mammalian proto-oncogene bcl-2 , 1994, Cell.

[10]  N. Thornberry,et al.  Apopain/CPP32 cleaves proteins that are essential for cellular repair: a fundamental principle of apoptotic death , 1996, The Journal of experimental medicine.

[11]  G M Bokoch,et al.  Membrane and morphological changes in apoptotic cells regulated by caspase-mediated activation of PAK2. , 1997, Science.

[12]  D. Vaux,et al.  The molecular biology of apoptosis. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[13]  David E. Housman,et al.  Hypoxia-mediated selection of cells with diminished apoptotic potential in solid tumours , 1996, Nature.

[14]  R. J. Clem,et al.  Prevention of apoptosis by a baculovirus gene during infection of insect cells. , 1991, Science.

[15]  D. Green,et al.  Cell‐free reconstitution of Fas‐, UV radiation‐ and ceramide‐induced apoptosis. , 1995, The EMBO journal.

[16]  M. Hengartner.  Programmed cell death in invertebrates. , 1996, Current opinion in genetics & development.

[17]  H. Horvitz,et al.  Programmed cell death in Caenorhabditis elegans. , 1994, Current opinion in genetics & development.

[18]  N. Thornberry,et al.  A Combinatorial Approach Defines Specificities of Members of the Caspase Family and Granzyme B , 1997, The Journal of Biological Chemistry.

[19]  H. Steller.  Mechanisms and genes of cellular suicide , 1995, Science.

[20]  S. Kaufmann,et al.  Chemotherapy-induced apoptosis. , 1997, Advances in pharmacology.

[21]  B. Vogelstein,et al.  A mammalian cell cycle checkpoint pathway utilizing p53 and GADD45 is defective in ataxia-telangiectasia , 1992, Cell.

[22]  Xiaodong Wang,et al.  Apaf-1, a Human Protein Homologous to C. elegans CED-4, Participates in Cytochrome c–Dependent Activation of Caspase-3 , 1997, Cell.

[23]  T. Mak,et al.  Stress-signalling kinase Sek1 protects thymocytes from apoptosis mediated by CD95 and CD3 , 1997, Nature.

[24]  C. Purdie,et al.  Thymocyte apoptosis induced by p53-dependent and independent pathways , 1993, Nature.

[25]  A. Levine,et al.  p53 and its 14 kDa C-terminal domain recognize primary DNA damage in the form of insertion/deletion mismatches , 1995, Cell.

[26]  H. Horvitz,et al.  The Caenorhabditis elegans cell-death protein CED-3 is a cysteine protease with substrate specificities similar to those of the human CPP32 protease. , 1996, Genes & development.

[27]  Patrick R. Griffin,et al.  Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis , 1995, Nature.

[28]  Junying Yuan,et al.  Human ICE/CED-3 Protease Nomenclature , 1996, Cell.

[29]  A. Chinnaiyan,et al.  Cytotoxic T-cell-derived granzyme B activates the apoptotic protease ICE-LAP3 , 1996, Current Biology.

[30]  T. Ley,et al.  Cleavage of CPP32 by Granzyme B Represents a Critical Role for Granzyme B in the Induction of Target Cell DNA Fragmentation* , 1996, The Journal of Biological Chemistry.

[31]  V. Fadok,et al.  Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. , 1992, Journal of immunology.

[32]  John Calvin Reed.  Double identity for proteins of the Bcl-2 family , 1997, Nature.

[33]  Jun O. Liu,et al.  p53-independent apoptosis induced by paclitaxel through an indirect mechanism. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[34]  C. Ware,et al.  A cysteine protease inhibitor prevents activation-induced T-cell apoptosis and death of peripheral blood cells from human immunodeficiency virus-infected individuals by inhibiting upregulation of Fas ligand. , 1997, Blood.

[35]  Xiaodong Wang,et al.  DFF, a Heterodimeric Protein That Functions Downstream of Caspase-3 to Trigger DNA Fragmentation during Apoptosis , 1997, Cell.

[36]  G. Salvesen,et al.  Proteolytic activation of the cell death protease Yama/CPP32 by granzyme B. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[37]  Shai Shaham,et al.  The C. elegans cell death gene ced-3 encodes a protein similar to mammalian interleukin-1β-converting enzyme , 1993, Cell.

[38]  E. White,et al.  Death-defying acts: a meeting review on apoptosis. , 1993, Genes & development.

[39]  E. Reinherz,et al.  T‐cell receptor ligation by peptide/MHC induces activation of a caspase in immature thymocytes: the molecular basis of negative selection , 1997, The EMBO journal.

[40]  M. Hayden,et al.  Cleavage of huntingtin by apopain, a proapoptotic cysteine protease, is modulated by the polyglutamine tract , 1996, Nature Genetics.

[41]  J. Giorgi,et al.  Sensitive method for measuring apoptosis and cell surface phenotype in human thymocytes by flow cytometry. , 1994, Cytometry.

[42]  N. Thornberry.  Interleukin-1 beta converting enzyme. , 1994, Methods in enzymology.

[43]  M. Cooke,et al.  Elimination of self-reactive B lymphocytes proceeds in two stages: Arrested development and cell death , 1993, Cell.

[44]  W. Liles.  Apoptosis – role in infection and inflammation , 1997 .

[45]  A. Culhane,et al.  Apoptosis: molecular regulation of cell death. , 1996, European journal of biochemistry.

[46]  S. Korsmeyer,et al.  bax-deficiency promotes drug resistance and oncogenic transformation by attenuating p53-dependent apoptosis. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[47]  Yuri Lazebnik,et al.  Multiple species of CPP32 and Mch2 are the major active caspases present in apoptotic cells , 1997, The EMBO journal.

[48]  H. Shinzawa,et al.  In vivo regulation of rat neutrophil apoptosis occurring spontaneously or induced with TNF-alpha or cycloheximide. , 1995, Journal of immunology.

[49]  Gwyn T. Williams,et al.  Antibodies to CD3/T-cell receptor complex induce death by apoptosis in immature T cells in thymic cultures , 1989, Nature.

[50]  C. Thompson,et al.  Apoptosis in the pathogenesis and treatment of disease , 1995, Science.

[51]  Muneesh Tewari,et al.  Yama/CPP32β, a mammalian homolog of CED-3, is a CrmA-inhibitable protease that cleaves the death substrate poly(ADP-ribose) polymerase , 1995, Cell.

[52]  W. Earnshaw,et al.  ICE-related proteases in apoptosis. , 1996, Current opinion in genetics & development.

[53]  Keisuke Kuida,et al.  Decreased apoptosis in the brain and premature lethality in CPP32-deficient mice , 1996, Nature.

[54]  S. Lowe,et al.  Selective induction of p53 and chemosensitivity in RB-deficient cells by E1A mutants unable to bind the RB-related proteins. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[55]  S. Lowe,et al.  Oncogenic ras Provokes Premature Cell Senescence Associated with Accumulation of p53 and p16INK4a , 1997, Cell.

[56]  E. Alnemri,et al.  CPP32, a novel human apoptotic protein with homology to Caenorhabditis elegans cell death protein Ced-3 and mammalian interleukin-1 beta-converting enzyme. , 1994, The Journal of biological chemistry.

[57]  D. Green,et al.  The killer and the executioner: how apoptosis controls malignancy. , 1995, Current opinion in immunology.