Induction of cell death by myristylated death domain of p55 TNF receptor is not abolished by Iprcg-like point mutation in death domain.
We transiently expressed the intracellular domains of p55 TNF receptor (TNFR1) as either a cytosolic- or a membrane-associated form and examined their effects on the endogenous receptor-mediated gene expression as well as on cell viability. We found that gene expression as measured by luciferase activity under NF-kappa B-controlling elements was blocked by all forms of the intracellular domain of TNFR1. The blockade of reporter gene expression was due to the cell death induced by the intracellular domain of TNFR1 per se. The killing mechanism of the intracellular domain peptides appeared to be apoptotic. Interestingly, myristylated form of the intracellular domain, consisting of mainly death domain showed the most potent cell-killing activity. Moreover, this myristylated death domain could still induce cell death even if lprcg-like point mutation (Leu351 to Ala), which has been reported to abrogate TNF-induced cytotoxicity, was introduced. This result suggests that the myristylated death domain activates an additional death signaling pathway which is not involved in TNF-induced cell death.
Kim
C H
CH
Biotech Research Institute, LG Chem Research Park, Yu-Song, Tae-Jeon, Korea.
Song
Y H
YH
Park
K
K
Oh
Y
Y
Lee
T H
TH
D017209:Apoptosis / Q000502:physiology*; D001483:Base Sequence; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism*; D015335:Molecular Probes / Q000235:genetics; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000502:physiology*; D017354:Point Mutation; D018124:Receptors, Tumor Necrosis Factor / Q000235:genetics* / Q000502:physiology*; D015398:Signal Transduction; D014407:Tumor Cells, Cultured
D015335:Molecular Probes; D010446:Peptide Fragments; D018124:Receptors, Tumor Necrosis Factor