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  • Title: Characterization of the rat urokinase plasminogen activator receptor promoter in PC12 cells.
    Author: Su F, Kozak KR, Herschman H, Reddy ST, Farias-Eisner R.
    Journal: J Neurosci Res; 2007 Jul; 85(9):1952-8. PubMed ID: 17455298.
    Abstract:
    Rat PC12 pheochromocytoma cells treated with nerve growth factor (NGF) extend "neurites" and initiate a neuronal differentiation pathway. Although neurotrophins, growth factors [e.g., epidermal growth factor (EGF)], and other ligands induce many common primary response genes (PRGs) in PC12 cells, a unique PRG subset is induced preferentially by NGF. Expression of one NGF preferentially induced gene, urokinase plasminogen activator receptor (UPAR), is required for NGF-induced neurite extension and neuronal differentiation. A 2.1-kb fragment of the rat UPAR 5' regulatory region confers differential expression by NGF versus EGF, following transfection of a luciferase reporter construct into PC12 cells. Deletion studies identified a region between -100 and -50 nucleotides from the transcription start site as the region conferring preferential NGF induction. Sequence comparisons among rat, human, and murine UPAR promoters identified two common potential regulatory regions. Site-directed mutation identified an activator protein-1 (AP-1) region between -66 and -72 bp, required for luciferase reporter activation by NGF. Electrophoretic mobility shift and antibody supershift assays demonstrated that specific Fos and Jun family members preferentially bind to this site following NGF treatment. We conclude that preferential activation of transcription factor binding at this AP-1 site mediates preferential NGF activation of the UPAR gene.
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