A Dominant Negative Fas-associated Death Domain Protein Mutant Inhibits Proliferation and Leads to Impaired Calcium Mobilization in Both T-cells and Fibroblasts

• Published in: The Journal of biological chemistry 2000-04, Vol.275 (14), p.10453-10462
• Main Authors: Hueber, Anne-Odile, Zörnig, Martin, Bernard, Anne-Marie, Chautan, Magali, Evan, Gerard
• Format: Article
• Language: English
• Subjects: 3T3 Cells; Adaptor Proteins, Signal Transducing; Animals; Apoptosis; Bombesin - pharmacology; Calcium - metabolism; Carrier Proteins - chemistry; Carrier Proteins - genetics; Carrier Proteins - physiology; caspase; Cell Cycle - physiology; Cell Division - drug effects; Fas antigen; Fas-Associated Death Domain Protein; FasL protein; Fibroblasts - cytology; Fibroblasts - physiology; Gene Expression Regulation - immunology; Interleukin-2 - genetics; Interleukin-6 - genetics; Ionomycin - pharmacology; Lck protein; Mice; Mice, Transgenic; Mutagenesis, Site-Directed; Promoter Regions, Genetic; Signal Transduction; Spleen - immunology; T-Lymphocytes - cytology; T-Lymphocytes - physiology; Transcription, Genetic
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.275.14.10453

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 G0→ S transition and in the G1 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.