Molecular Cloning and Characterization of the Human <i>VGF</i> Promoter Region

Abstract

The VGF gene encodes a secretory protein that is expressed in a cell type-restricted pattern in neuroendocrine cells and is up-regulated by nerve growth factor (NGF) in the rat pheochromocytoma PC12 cell line. Here we report the isolation and characterization of the 5'-terminal region of the human VGF gene. In addition to a TATA box and a CCAAT box located at canonical distances from the transcription start site, the human VGF promoter contains several consensus sequences for different transcription factors, including a cyclic AMP response element and an AP-1 element, several GC boxes, and sequences homologous to other neuronal promoters. Transient transfection analysis demonstrates that 2.3 kb of the 5'-flanking sequence acts as a tissue-specific promoter, efficiently used only by neuronal cells that express endogenous VGF. Deletion analysis reveals that a positive regulatory region is located between nucleotides -458 to -204. Negative cis-acting elements that repress promoter activity in cell lines that do not normally express VGF are located between nucleotides -2,305 and -573 and between -458 and -204. The 5'-flanking region of the human VGF gene confers responsiveness to NGF, cyclic AMP, and phorbol ester treatment.