A dominant negative Fas-associated death domain protein mutant inhibits proliferation and leads to impaired calcium mobilization in both T-cells and fibroblasts

J Biol Chem. 2000 Apr 7;275(14):10453-62. doi: 10.1074/jbc.275.14.10453.

Abstract

Death domain-containing members of the tumor necrosis factor (TNF) receptor family ("death receptors") can induce apoptosis upon stimulation by their natural ligands or by agonistic antibodies. Activated death receptors recruit death domain adapter proteins like Fas-associated death domain protein (FADD), and this ultimately leads to proteolytic activation of the caspase cascade and cell death. Recently, FADD has also been implicated in the regulation of proliferation; functional inhibition of FADD results in p53-dependent impairment of proliferation in activated T-cells. In this study we have further analyzed T-cells derived from transgenic mice expressing a dominant negative FADD mutant (FADD DN) under control of the lck promoter in vitro so as to identify the signaling pathways that become engaged upon T-cell receptor stimulation and that are regulated by death receptors. FADD DN expression inhibits T-cell proliferation, both at the G(0) --> S transition and in the G(1) phase of continuously proliferating cells. We observe a decrease in the release of calcium from intracellular stores after T-cell receptor stimulation, whereas influx of extracellular calcium seems to be unaffected. FADD DN-expressing fibroblasts show a similarly inhibited cell growth and impaired calcium mobilization indicating that the modulation of proliferation and calcium response by death receptors is not cell type-specific.

MeSH terms

  • 3T3 Cells
  • Adaptor Proteins, Signal Transducing*
  • Animals
  • Apoptosis
  • Bombesin / pharmacology
  • Calcium / metabolism*
  • Carrier Proteins / chemistry
  • Carrier Proteins / genetics
  • Carrier Proteins / physiology*
  • Cell Cycle / physiology*
  • Cell Division / drug effects
  • Fas-Associated Death Domain Protein
  • Fibroblasts / cytology
  • Fibroblasts / physiology
  • Gene Expression Regulation / immunology
  • Interleukin-2 / genetics
  • Interleukin-6 / genetics
  • Ionomycin / pharmacology
  • Mice
  • Mice, Transgenic
  • Mutagenesis, Site-Directed
  • Promoter Regions, Genetic
  • Signal Transduction
  • Spleen / immunology
  • T-Lymphocytes / cytology
  • T-Lymphocytes / physiology*
  • Transcription, Genetic

Substances

  • Adaptor Proteins, Signal Transducing
  • Carrier Proteins
  • Fadd protein, mouse
  • Fas-Associated Death Domain Protein
  • Interleukin-2
  • Interleukin-6
  • Ionomycin
  • Bombesin
  • Calcium