TY - JOUR
T1 - Lipopolysaccharide-inducible and salicylate-sensitive nuclear factor(s) on human tumor necrosis factor alpha promoter
AU - Takashiba, S.
AU - Van Dyke, T. E.
AU - Shapira, L.
AU - Amar, S.
PY - 1995
Y1 - 1995
N2 - Lipopolysaccharide (LPS) is one of the most potent trigger substances for monocytes and macrophages causing secretion of tumor necrosis factor alpha (TNF-α) and other inflammatory mediators. The nature of the nuclear factors involved in human TNF-α gene regulation is still unknown. Nuclear factor kappa B (NF-κB) proteins have been suggested to play an important role in gene transcription of inflammatory mediators when monocytes are stimulated with LPS. However, it remains unclear whether these nuclear factors are the only ones involved in human TNF-α gene regulation. In this report, to further the identification of nuclear factor(s) involved in TNF-α gene regulation, human monocytic THP-1 cells were transfected with a series of truncated versions of human TNF-α promoter. A 98-bp region located from nucleotides -584 to -487 demonstrated strong promoter activity. Electrophoretic mobility shift assays demonstrated that a 64-bp fragment located within the 98-bp region and lacking any potential NF-κB-binding sites avidly bound LPS-challenged THP-1 nuclear protein. Although this binding was inhibited in salicylate-treated cells, as was binding of NF-κB, the pattern of binding was found to differ from that noted for NF-κB. Analysis of this 64-bp fragment disclosed the absence of an NF-κB consensus sequence, suggesting a novel nuclear DNA-binding protein necessary for the initiation of human TNF-α transcription other than, or in addition to, NF- κB.
AB - Lipopolysaccharide (LPS) is one of the most potent trigger substances for monocytes and macrophages causing secretion of tumor necrosis factor alpha (TNF-α) and other inflammatory mediators. The nature of the nuclear factors involved in human TNF-α gene regulation is still unknown. Nuclear factor kappa B (NF-κB) proteins have been suggested to play an important role in gene transcription of inflammatory mediators when monocytes are stimulated with LPS. However, it remains unclear whether these nuclear factors are the only ones involved in human TNF-α gene regulation. In this report, to further the identification of nuclear factor(s) involved in TNF-α gene regulation, human monocytic THP-1 cells were transfected with a series of truncated versions of human TNF-α promoter. A 98-bp region located from nucleotides -584 to -487 demonstrated strong promoter activity. Electrophoretic mobility shift assays demonstrated that a 64-bp fragment located within the 98-bp region and lacking any potential NF-κB-binding sites avidly bound LPS-challenged THP-1 nuclear protein. Although this binding was inhibited in salicylate-treated cells, as was binding of NF-κB, the pattern of binding was found to differ from that noted for NF-κB. Analysis of this 64-bp fragment disclosed the absence of an NF-κB consensus sequence, suggesting a novel nuclear DNA-binding protein necessary for the initiation of human TNF-α transcription other than, or in addition to, NF- κB.
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U2 - 10.1128/iai.63.4.1529-1534.1995
DO - 10.1128/iai.63.4.1529-1534.1995
M3 - Article
C2 - 7890420
AN - SCOPUS:0028937836
VL - 63
SP - 1529
EP - 1534
JO - Infection and Immunity
JF - Infection and Immunity
SN - 0019-9567
IS - 4
ER -