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• W4313343417 endingPage "4234" @default.
• W4313343417 startingPage "4226" @default.
• W4313343417 abstract "Abstract Among its plethora of activities as an inflammatory mediator, TNF-α has potent regulatory control on extracellular matrix production and degradation. Earlier studies have documented that TNF-α inhibits type I collagen gene (COL1A2) expression at the transcriptional level, but the characterization of the transcription factors involved has been elusive. In the present study, using transient cell transfection of human dermal fibroblasts with a battery of 5′ end deletion/chloramphenicol acetyltransferase (CAT) reporter gene constructs, we have characterized the TNF-α response element of the COL1A2 promoter. The TNF-α response element was attributed to a specific region that comprises noncanonical activator protein-1 (AP-1) (CGAGTCA) and NF-κB (AGAGTTTCCC) binding sites. TNF-α effect was eliminated by a 2-bp substitution mutation in the NF-κB1 binding half site of the NF-κB cis element. Electrophoretic mobility shift assays (EMSA) showed that recombinant human NF-κB heterodimers as well as NF-κB1 and RelA homodimers, but not AP-1, were capable of binding this element. Further, EMSA with human fibroblast nuclear extracts demonstrated enhanced binding of a single, specific complex within 5 min of TNF-α stimulation, which reached a plateau by 1 h and was not affected by preincubation of cells with cycloheximide. Gel supershift assays identified the complex as the NF-κB (p50/p65) heterodimer, whereas Abs to nuclear factor of activated T cells (NF-AT) and Jun family members failed to recognize the complex. These data suggest that in fibroblasts TNF-α activates and initiates the nuclear translocation of NF-κB that binds a divergent NF-κB element and plays a critical role in the observed inhibition of α2(I) collagen gene transcription." @default.
• W4313343417 created "2023-01-06" @default.
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• W4313343417 date "1999-04-01" @default.
• W4313343417 modified "2023-10-17" @default.
• W4313343417 title "Nuclear Factor-κB Mediates TNF-α Inhibitory Effect on α2(I) Collagen (COL1A2) Gene Transcription in Human Dermal Fibroblasts" @default.
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• W4313343417 doi "https://doi.org/10.4049/jimmunol.162.7.4226" @default.
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