TNF induction of jagged-1 in endothelial cells is NFkappaB-dependent.
TNF-alpha is a potent proinflammatory cytokine that induces endothelial cell (EC) adhesion molecules. In addition, TNF promotes angiogenesis by inducing an EC tip cell phenotype and the expression of jagged-1, a ligand for the notch pathway. Notch signaling is critical for vascular patterning and helps to restrict the proliferation of tip cells. Here we demonstrate that TNF induction of jagged-1 in human EC is rapid and dependent upon signaling through TNFR1, but not TNFR2. A luciferase reporter construct carrying 3.7 kb of 5' promoter sequence from the human gene was responsive to both TNF and overexpression of NFkappaB pathway components. TNF-induced promoter activation was blocked by treatment with an NFkappaB inhibitor or co-expression of dominant-negative IKKbeta. Mutations in a putative NFkappaB-binding site at -3.0 kb, which is conserved across multiple species, resulted in a loss of responsiveness to TNF and NFkappaB. Electromobility shift and chromatin immunoprecipitation assays revealed binding of both p50 and p65 to the promoter in response to TNF treatment. Full promoter activity also depends on an AP-1 site at -2.0 kb. These results indicate that canonical NFkappaB signaling is required for TNF induction of the notch ligand jagged-1 in EC.
Johnston
Douglas A
DA
Department of Molecular Biology and Biochemistry, University of California Irvine, 92697, USA.
Dong
Bamboo
B
Hughes
Christopher C W
CC
D001483:Base Sequence; D001665:Binding Sites; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000378:metabolism*; D005434:Flow Cytometry; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D000072100:Jagged-1 Protein; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000378:metabolism; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000378:metabolism; D000072099:Serrate-Jagged Proteins; D015398:Signal Transduction; D018808:Transcription Factor AP-1 / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
D002135:Calcium-Binding Proteins; D036341:Intercellular Signaling Peptides and Proteins; C000606273:JAG1 protein, human; D000072100:Jagged-1 Protein; D008565:Membrane Proteins; D016328:NF-kappa B; D047888:Receptors, Tumor Necrosis Factor, Type I; D047889:Receptors, Tumor Necrosis Factor, Type II; D000072099:Serrate-Jagged Proteins; D018808:Transcription Factor AP-1; D014409:Tumor Necrosis Factor-alpha