Apoptotic crosstalk of TNF receptors: TNF-R2-induces depletion of TRAF2 and IAP proteins and accelerates TNF-R1-dependent activation of caspase-8.

We have recently shown that stimulation of TNF-R2 selectively enhances apoptosis induction by the death receptor TNF-R1. Here, we demonstrate that stimulation of CD30 or CD40 also leads to selective enhancement of TNF-R1-induced cell death. Enhancement of apoptosis was correlated with the depletion of endogenous TRAF2 within 1 to 6 hours. Selective prestimulation of TNF-R2 for several hours inhibited TNF-R2-induced activation of the anti-apoptotic NF-kappaB pathway up to 90% and dramatically enhanced apoptosis induction by this receptor. When both TNF-receptors were stimulated simultaneously, TNF-R1-induced NF-kappaB activation remained unaffected but TNF-R1-induced apoptosis was still significantly enhanced. Compared with FasL-induced cell death TNF-R1-induced activation of caspase-8 was significantly weaker and delayed. Costimulation or prestimulation of TNF-R2 enhanced caspase-8 processing. Life cell imaging and confocal microscopy revealed that both TNF-R1 and TNF-R2 recruited the anti-apoptotic factor cIAP1 in a TRAF2-dependent manner. Thus, TNF-R2 may compete with TNF-R1 for the recruitment of newly synthesized TRAF2-bound anti-apoptotic factors, thereby promoting the formation of a caspase-8-activating TNF-R1 complex. Hence, TNF-R2 triggering can interfere with TNF-R1-induced apoptosis by inhibition of NF-kappaB-dependent production of anti-apoptotic factors and by blocking the action of anti-apoptotic factors at the post-transcriptional level.

[1]  Peter Scheurich,et al.  Induction of cell death by tumour necrosis factor (TNF) receptor 2, CD40 and CD30: a role for TNF‐R1 activation by endogenous membrane‐anchored TNF , 1999, The EMBO journal.

[2]  P. Scheurich,et al.  Enhancement of TNF receptor p60-mediated cytotoxicity by TNF receptor p80: requirement of the TNF receptor-associated factor-2 binding site. , 1997, Journal of immunology.

[3]  K. Brown,et al.  Differential Signaling and Tumor Necrosis Factor Receptor–Associated Factor (Traf) Degradation Mediated by Cd40 and the Epstein-Barr Virus Oncoprotein Latent Membrane Protein 1 (Lmp1) , 2001, The Journal of experimental medicine.

[4]  Peter Scheurich,et al.  NF-κB Inducers Upregulate cFLIP, a Cycloheximide-Sensitive Inhibitor of Death Receptor Signaling , 2001, Molecular and Cellular Biology.

[5]  E. Bröcker,et al.  Regulation of tumor necrosis factor-related apoptosis-inducing ligand sensitivity in primary and transformed human keratinocytes. , 2000, Cancer research.

[6]  T. Mak,et al.  Critical Roles of TRAF2 and TRAF5 in Tumor Necrosis Factor-induced NF-κB Activation and Protection from Cell Death* , 2001, The Journal of Biological Chemistry.

[7]  Hong-Bing Shu,et al.  TRADD–TRAF2 and TRADD–FADD Interactions Define Two Distinct TNF Receptor 1 Signal Transduction Pathways , 1996, Cell.

[8]  M. Malim,et al.  Suppression of tumor necrosis factor-induced cell death by inhibitor of apoptosis c-IAP2 is under NF-kappaB control. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[9]  C. Thompson,et al.  Translocation of TRAF proteins regulates apoptotic threshold of cells. , 2000, Biochemical and biophysical research communications.

[10]  Hiroshi Takayanagi,et al.  T-cell-mediated regulation of osteoclastogenesis by signalling cross-talk between RANKL and IFN-γ , 2000, Nature.

[11]  C. Duckett,et al.  Differential effects of CD30 activation in anaplastic large cell lymphoma and Hodgkin disease cells. , 2000, Blood.

[12]  Mike Rothe,et al.  Tumor necrosis factor's cytotoxic activity is signaled by the p55 TNF receptor , 1993, Cell.

[13]  P. Scheurich,et al.  TNFR80-dependent enhancement of TNFR60-induced cell death is mediated by TNFR-associated factor 2 and is specific for TNFR60. , 1998, Journal of immunology.

[14]  George Kollias,et al.  The transmembrane form of tumor necrosis factor is the prime activating ligand of the 80 kDa tumor necrosis factor receptor , 1995, Cell.

[15]  W. Fiers,et al.  Cooperation of both TNF receptors in inducing apoptosis: involvement of the TNF receptor-associated factor binding domain of the TNF receptor 75. , 1998, Journal of immunology.

[16]  Klaus Rajewsky,et al.  NEMO/IKKγ-Deficient Mice Model Incontinentia Pigmenti , 2000 .

[17]  C. Thompson,et al.  CD30-dependent degradation of TRAF2: implications for negative regulation of TRAF signaling and the control of cell survival. , 1997, Genes & development.

[18]  D. Goeddel,et al.  Embryonic Lethality, Liver Degeneration, and Impaired NF-κB Activation in IKK-β-Deficient Mice , 1999 .

[19]  I. Beletsky,et al.  Dual role of the p75 tumor necrosis factor (TNF) receptor in TNF cytotoxicity , 1994, The Journal of experimental medicine.

[20]  M. Kelliher,et al.  The distinct roles of TRAF2 and RIP in IKK activation by TNF-R1: TRAF2 recruits IKK to TNF-R1 while RIP mediates IKK activation. , 2000, Immunity.

[21]  Stefan Grimm,et al.  The Death Domain Kinase RIP Mediates the TNF-Induced NF-κB Signal , 1998 .

[22]  J. Tschopp,et al.  Caspase‐induced inactivation of the anti‐apoptotic TRAF1 during Fas ligand‐mediated apoptosis , 2000, FEBS letters.

[23]  Q Cheng,et al.  NF-kappaB-mediated up-regulation of Bcl-x and Bfl-1/A1 is required for CD40 survival signaling in B lymphocytes. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[24]  K. Frei,et al.  Severity of symptoms and demyelination in MOG‐induced EAE depends on TNFR1 , 1999, European journal of immunology.

[25]  M. Lenardo,et al.  Induction of apoptosis in mature T cells by tumour necrosis factor , 1995, Nature.

[26]  W. Fiers,et al.  Both TNF receptors are required for TNF-mediated induction of apoptosis in PC60 cells. , 1995, Journal of immunology.

[27]  P. Scheurich,et al.  Inhibition of Death Receptor-mediated Gene Induction by a Cycloheximide-sensitive Factor Occurs at the Level of or Upstream of Fas-associated Death Domain Protein (FADD)* , 2000, The Journal of Biological Chemistry.

[28]  David Baltimore,et al.  Embryonic lethality and liver degeneration in mice lacking the RelA component of NF-κB , 1995, Nature.

[29]  T. Griffith,et al.  Intracellular regulation of TRAIL-induced apoptosis in human melanoma cells. , 1998, Journal of immunology.

[30]  J. Tschopp,et al.  Conversion of Membrane-bound Fas(CD95) Ligand to Its Soluble Form Is Associated with Downregulation of Its Proapoptotic Activity and Loss of Liver Toxicity , 1998, The Journal of experimental medicine.

[31]  P. Scheurich,et al.  TNF receptor associated factors in cytokine signaling. , 1999, Cytokine & growth factor reviews.

[32]  Hong-Hee Kim,et al.  Caspase‐mediated cleavage of TRAF3 in FasL‐stimulated Jurkat‐T cells , 2001, Journal of leukocyte biology.

[33]  Mike Rothe,et al.  The TNFR2-TRAF signaling complex contains two novel proteins related to baculoviral inhibitor of apoptosis proteins , 1995, Cell.

[34]  Inder M. Verma,et al.  Severe Liver Degeneration in Mice Lacking the IκB Kinase 2 Gene , 1999 .

[35]  W. Liao,et al.  The essential role of MEKK3 in TNF-induced NF-κB activation , 2001, Nature Immunology.

[36]  Y. Goltsev,et al.  Tumor necrosis factor receptor and Fas signaling mechanisms. , 1999, Annual review of immunology.

[37]  John Calvin Reed,et al.  The c‐IAP‐1 and c‐IAP‐2 proteins are direct inhibitors of specific caspases , 1997, The EMBO journal.

[38]  Matthias Mann,et al.  FLICE is activated by association with the CD95 death‐inducing signaling complex (DISC) , 1997, The EMBO journal.

[39]  P. Scheurich,et al.  TR60 and TR80 tumor necrosis factor (TNF)-receptors can independently mediate cytolysis. , 1993, Lymphokine and cytokine research.

[40]  D. Goeddel,et al.  The tumor necrosis factor receptor 2 signal transducers TRAF2 and c-IAP1 are components of the tumor necrosis factor receptor 1 signaling complex. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[41]  B. Seed,et al.  Regulated commitment of TNF receptor signaling: a molecular switch for death or activation. , 1999, Immunity.

[42]  W. Fiers,et al.  Cytotoxicity in L929 murine fibrosarcoma cells after triggering of transfected human p75 tumour necrosis factor (TNF) receptor is mediated by endogenous murine TNF. , 1995, Cytokine.

[43]  N. Fan,et al.  The p70 tumor necrosis factor receptor mediates cytotoxicity , 1992, Cell.

[44]  P. Scheurich,et al.  The TNF-receptor-associated factor family: scaffold molecules for cytokine receptors, kinases and their regulators. , 2001, Cellular signalling.

[45]  V. Godfrey,et al.  Female mice heterozygous for IKK gamma/NEMO deficiencies develop a dermatopathy similar to the human X-linked disorder incontinentia pigmenti. , 2000, Molecular cell.

[46]  M. Karin,et al.  The α and β Subunits of IκB Kinase (IKK) Mediate TRAF2-Dependent IKK Recruitment to Tumor Necrosis Factor (TNF) Receptor 1 in Response to TNF , 2001, Molecular and Cellular Biology.

[47]  M. Lenardo,et al.  A crucial role for p80 TNF‐R2 in amplifying p60 TNF‐R1 apoptosis signals in T lymphocytes , 2000, European journal of immunology.

[48]  T. Espevik,et al.  Involvement of the tumor necrosis factor receptor p75 in mediating cytotoxicity and gene regulating activities , 1994, European journal of immunology.

[49]  R. Locksley,et al.  The TNF and TNF Receptor Superfamilies Integrating Mammalian Biology , 2001, Cell.

[50]  P. Henkart,et al.  Cytotoxic effect of TNF and lymphotoxin on T lymphoblasts. , 1995, Journal of immunology.

[51]  Peter Scheurich,et al.  The Human Tumor Necrosis Factor (TNF) Receptor-associated Factor 1 Gene (TRAF1) Is Up-regulated by Cytokines of the TNF Ligand Family and Modulates TNF-induced Activation of NF-κB and c-Jun N-terminal Kinase* , 1999, The Journal of Biological Chemistry.

[52]  H. Engelmann,et al.  A novel function of CD40: induction of cell death in transformed cells , 1996, The Journal of experimental medicine.

[53]  P. Scheurich,et al.  The type 1 receptor (CD120a) is the high-affinity receptor for soluble tumor necrosis factor. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[54]  T. Mak,et al.  Tumor necrosis factor receptor p55 mediates deletion of peripheral cytotoxic T lymphocytes in vivo , 1996, European journal of immunology.

[55]  T. Mak,et al.  Severe liver degeneration and lack of NF-kappaB activation in NEMO/IKKgamma-deficient mice. , 2000, Genes & development.

[56]  P. Scheurich,et al.  Dominant-negative FADD inhibits TNFR60-, Fas/Apo1- and TRAIL-R/Apo2-mediated cell death but not gene induction , 1998, Current Biology.

[57]  C. Y. Wang,et al.  NF-kappaB antiapoptosis: induction of TRAF1 and TRAF2 and c-IAP1 and c-IAP2 to suppress caspase-8 activation. , 1998, Science.