Sequences required for induction of neurotensin receptor gene expression during neuronal differentiation of N1E-115 neuroblastoma cells
Student AuthorsKeith Tully
UMass Chan AffiliationsDepartment of Molecular Genetics and Microbiology
Graduate School of Biomedical Sciences
Document TypeJournal Article
KeywordsAnimals; Base Sequence; *Cell Differentiation; Cloning, Molecular; DNA; Dimethyl Sulfoxide; Gene Expression Regulation; Mice; Molecular Sequence Data; Neuroblastoma; Neurons; Promoter Regions (Genetics); Receptors, Neurotensin; Regulatory Sequences, Nucleic Acid; Sequence Homology, Nucleic Acid; Tumor Cells, Cultured
Medicine and Health Sciences
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AbstractThe promoter region of the mouse high affinity neurotensin receptor (Ntr-1) gene was characterized, and sequences required for expression in neuroblastoma cell lines that express high affinity NT-binding sites were characterized. Me(2)SO-induced neuronal differentiation of N1E-115 neuroblastoma cells increased both the expression of the endogenous Ntr-1 gene and reporter genes driven by NTR-1 promoter sequences by 3-4-fold. Deletion analysis revealed that an 83-base pair promoter region containing the transcriptional start site is required for Me(2)SO activation. Detailed mutational analysis of this region revealed that a CACCC box and the central region of a large GC-rich palindrome are the crucial cis-regulatory elements required for Me(2)SO induction. The CACCC box is bound by at least one factor that is induced upon Me(2)SO treatment of N1E-115 cells. The Me(2)SO effect was found to be both selective and cell type-restricted. Basal expression in the neuroblastoma cell lines required a distinct set of sequences, including an Sp1-like sequence, and a sequence resembling an NGFI-A-binding site; however, a more distal 5' sequence was found to repress basal activity in N1E-115 cells. These results provide evidence that Ntr-1 gene regulation involves both positive and negative regulatory elements located in the 5'-flanking region and that Ntr-1 gene activation involves the coordinate activation or induction of several factors, including a CACCC box binding complex.
J Biol Chem. 1999 Oct 15;274(42):30066-79.
Permanent Link to this Itemhttp://hdl.handle.net/20.500.14038/32673
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