Regulation of tumor necrosis factor alpha transcription in macrophages: involvement of four kappa B-like motifs and of constitutive and inducible forms of NF-kappa B.

摘要: This study characterizes the interaction of murine macrophage nuclear proteins with tumor necrosis factor alpha (TNF-alpha) promoter. Gel retardation and methylation interference assays showed that stimulation TNF-alpha gene transcription in peritoneal exudate macrophages was accompanied by induction DNA-binding recognized different affinities four elements related to kappa B consensus motif a Y-box motif. We suggest basal level expression is due binding constitutive form NF-kappa B, present at low levels nuclei from resting thioglycolate macrophages, some if not all motifs; we postulate this contains only 50-kilodalton (kDa) protein subunits 65-kDa (P. Baeuerle D. Baltimore, Genes Dev. 3:1689-1698, 1989). Agents such as glucocorticoids, which decrease transcription, diminished B. Stimulation lipopolysaccharide, gamma interferon, or cycloheximide led an increased content induced represents since it generated protein-DNA complexes slower mobility; propose both 50- subunits, latter ones being necessary bind its cytoplasmic inhibitor uninduced cells (Baeuerle Dev., In cells, inducible indeed detectable cytosol after deoxycholate treatment. UV cross-linking experiments gel indicated higher-order complex other proteins.