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• W2014726611 abstract "Plasminogen-activator inhibitor type 2 (PAI-2), a serine protease inhibitor involved in the regulation of urokinase-dependent proteolysis, is also implicated in the inhibition of tumor-necrosis-factor-(TNF)-mediated apoptosis. The PAI-2 gene is one of the most TNF-responsive genes known and is also highly induced by the phorbol ester phorbol 12-myristate 13-acetate (PMA) and the phosphatase inhibitor, okadaic acid, in both HT-1080 fibrosarcoma and U-937 histiocytic cells. We sought to identify and characterize regulatory cis-acting DNA elements and trans -acting factors which mediate basal and inducible PAI-2 gene transcription. A series of promoter deletion mutants (nucleotides –1859 to –91) fused to the chloramphenicol acetyl transferase (CAT) reporter gene were transfected into HT-1080 cells. Two repressor regions were identified; one distally between positions –1859 and –1100, and one proximally between positions –259 and –219. Cells transfected with constructs harboring more than 259 bp promoter sequence produced a 10–15-fold increase in CAT activity when treated with PMA or okadaic acid, but produced only a minimal (2.5-fold) increase in response to TNF. Removal of the proximal repressor by deletion to position –219, or by internal deletion from the –1100 PAI-2 CAT construct, resulted in a selective increase in TNF responsiveness, suggesting that induction of PAI-2 gene transcription by TNF is associated with derepression. Detailed analysis of the proximal repressor utilizing the electrophoretic mobility shift assay (EMSA), identified two novel and distinct protein-binding sites (A and B). Site A is located within the 40-bp proximal repressor while site B is situated immediately adjacent to the 3′ boundary. Treatment of cells with PMA or okadaic acid produced no change in the binding activity of proteins recognising sites A or B. However, treatment of cells with TNF results in a profound selective reduction in site-B-binding activity, suggesting that this site plays a significant role in TNF-mediated regulation of PAI-2 gene expression. Our findings suggest that TNF-mediated induction of PAI-2 gene expression involves derepression and is associated with cis -acting and trans -acting factors located within and adjacent to the proximal repressor region." @default.
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• W2014726611 date "1996-10-01" @default.
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• W2014726611 title "Molecular Mechanisms Governing Tumor-Necrosis-Factor-Mediated Regulation of Plasminogen-Activator Inhibitor Type-2 Gene Expression" @default.
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• W2014726611 doi "https://doi.org/10.1111/j.1432-1033.1996.0093t.x" @default.
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