title: Soluble TNF Regulates TACE via AP-2α Transcription Factor in Mouse Dendritic Cells.
abstract: Dendritic cells (DCs), the essential immunoregulatory and APCs, are major producers of the central mediator of inflammation, soluble TNF-α (sTNF). sTNF is generated by TNF-α converting enzyme (TACE) proteolytic release of the transmembrane TNF (tmTNF) ectodomain. The mechanisms of TACE and sTNF regulation in DCs remain elusive. This study newly defines that sTNF regulates TACE in mouse DCs by engaging the AP-2α transcription factor. We found that the expression of AP-2α was higher, whereas the expression and activity of TACE were lower, in wild-type DCs (wtDCs) than in TNF knockout (TNFko) DCs. Exogenous sTNF rapidly and simultaneously induced increases of AP-2α expression and decreases of TACE expression and activity in wtDCs and TNFko DCs, indicating that AP-2α and TACE are inversely dependent on sTNF and are functionally associated. To define this functional association, we identified an AP-2α binding site in TACE promoter and demonstrated, using EMSAs and chromatin immunoprecipitation assays, that AP-2α could bind to TACE promoter in a TNF-dependent manner. Additionally, sTNF simultaneously enhanced AP-2α expression and decreased TACE promoter luciferase activity in DCs. Similarly, transfection of AP-2α cDNA decreased TACE promoter luciferase activity, TACE expression, and TACE enzymatic activity in wtDCs or TNFko DCs. In contrast, transfection of AP-2α small interfering RNA increased TACE promoter luciferase activity, TACE expression, and TACE enzymatic activity in wtDCs. These results show that TACE is a target of, and is downregulated by, sTNF-induced AP-2α transcription factor in DCs.
authors.affiliation: Department of Pathology, University of Pittsburgh Cancer Institute, Pittsburgh, PA 15232.
authors.forename: Lisheng
authors.initials: L
authors.lastname: Ge
authors.affiliation: Department of Pathology, University of Pittsburgh Cancer Institute, Pittsburgh, PA 15232; vujanovicnl@upmc.edu.|Department of Immunology, University of Pittsburgh Cancer Institute, Pittsburgh, PA 15232; and.|VA Pittsburgh Healthcare System, Pittsburgh, PA 15261.
authors.forename: Nikola L
authors.initials: NL
authors.lastname: Vujanovic
mesh_terms: D000072198:ADAM17 Protein / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D047369:Chromatin Immunoprecipitation; D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005786:Gene Expression Regulation / Q000276:immunology; D015151:Immunoblotting; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D050656:Transcription Factor AP-2 / Q000276:immunology / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology / Q000378:metabolism*
chemical_list: D050656:Transcription Factor AP-2; D014409:Tumor Necrosis Factor-alpha; D000072198:ADAM17 Protein; C000606463:Adam17 protein, mouse