Isolation and characterization of the human secretogranin II gene promoter

• Published in: Brain research. Molecular brain research. 2000-01, Vol.75 (1), p.8-15
• Main Authors: Scammell, Jonathan G, Reddy, Shyla, Valentine, Donna L, Coker, Tracy N, Nikolopoulos, Sotiris N, Ross, Robert A
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Biological and medical sciences; Chromogranins; Cloning, Molecular; Consensus Sequence; Cyclic AMP response element-binding protein; Fundamental and applied biological sciences. Psychology; Granin; Humans; Luciferase reporter gene, neuroblastoma; Luciferases - genetics; Mice; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; PC12 Cells; Promoter Regions, Genetic; Protein Biosynthesis; Proteins - chemistry; Proteins - genetics; Rats; Recombinant Fusion Proteins - biosynthesis; secretogranin II; Sequence Alignment; Sequence Homology, Nucleic Acid; SgII gene; TATA Box; Transcription. Transcription factor. Splicing. Rna processing; Transfection; Tumor Cells, Cultured
• ISSN: 0169-328X; 1872-6941
• DOI: 10.1016/S0169-328X(99)00269-7

The goal of this study was to isolate and functionally characterize the human secretogranin II (SgII) gene promoter. SgII is a member of the granin family of proteins which are selectively expressed in neurosecretory cells. The human SgII promoter contains a consensus TATA box and cyclic AMP response element (CRE) 35 and 74 bp upstream of the transcription start site, respectively, elements also found in the mouse and rat SgII gene promoters. Transfection studies showed that 869 bp of the human SgII promoter were sufficient to confer cell type-specific expression of an SgII promoter-luciferase reporter gene in neurosecretory PC-12, GH and BE(2)-M17 cells. The activity of the human SgII promoter was also compared in three N-type, human neuroblastoma cell lines [BE(2)-M17, SMS-KAN and SH-SY5Y], which differ markedly in the level of SgII expression. SgII promoter activities in the neuroblastoma cell lines correlated not only with the levels of SgII but also the levels of the cyclic AMP response element-binding protein CREB which were highest in BE(2)-M17 cells and lowest in SH-SY5Y cells. To establish that the activity of the human SgII promoter in these neuroblastoma cell lines is dependent on the level of CREB, rat CREB was overexpressed in SH-SY5Y cells. SgII promoter activity was up to 8-fold higher in SH-SY5Y cells overexpressing CREB. These results suggest that SgII expression is a marker for neuronal differentiation in human neuroblastoma cell lines and is dependent on the level of CREB expression.