Molecular cloning and characterization of the human VGF promoter region.
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.
Canu
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Dipartimento di Medicina Sperimentale e Scienze Biomediche, Seconda Università di Roma Tor Vergata, Italy.
Possenti
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R
Rinaldi
A M
AM
Trani
E
E
Levi
A
A
D000818:Animals; D001483:Base Sequence; D002273:Carcinogens / Q000494:pharmacology; D003001:Cloning, Molecular; D016384:Consensus Sequence / Q000235:genetics; D000242:Cyclic AMP / Q000494:pharmacology; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects / Q000502:physiology; D017930:Genes, Reporter; D006801:Humans; D008969:Molecular Sequence Data; D009479:Neuropeptides; D016716:PC12 Cells / Q000502:physiology; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011506:Proteins / Q000235:genetics*; D051381:Rats; D017422:Sequence Analysis, DNA; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic / Q000235:genetics
D002273:Carcinogens; D009479:Neuropeptides; D011506:Proteins; C062040:Vgf protein, rat; D000242:Cyclic AMP; D013755:Tetradecanoylphorbol Acetate