Cytosolic calcium is essential in the basal expression of tyrosine hydroxylase gene.
To test the role of cytosolic calcium in the basal expression of a neuronal gene, promoter activity of the rat tyrosine hydroxylase (TH) gene was monitored upon reduced resting level of intracellular calcium. TH promoter activity was decreased by cell-permeable calcium chelator, BAPTA/AM, in SK-N-BE(2)C human neuroblastoma cells. The cAMP response element (CRE) was mapped to the calcium responsible element by mutational and deletional analysis of the 5' upstream promoter region. Gel shift assay showed 2 CRE-specific DNA-protein complexes. The quantities of specific complexes were markedly decreased in BAPTA/AM-treated cells. These data suggest that resting level of intracellular calcium has a critical role in the basal expression of TH gene through the regulation of the binding of nuclear proteins to the CRE motif in the promoter.
Chae
H D
HD
Department of Life Science, Pohang University of Science and Technology, Republic of Korea.
Kim
K T
KT
D000818:Animals; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002614:Chelating Agents / Q000494:pharmacology; D003600:Cytosol / Q000201:enzymology; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D015870:Gene Expression; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009447:Neuroblastoma; D009838:Oligodeoxyribonucleotides; D011401:Promoter Regions, Genetic / Q000187:drug effects; D051381:Rats; D017384:Sequence Deletion; D014407:Tumor Cells, Cultured; D014446:Tyrosine 3-Monooxygenase / Q000096:biosynthesis*
D002614:Chelating Agents; D009838:Oligodeoxyribonucleotides; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D004533:Egtazic Acid; D014446:Tyrosine 3-Monooxygenase; D002118:Calcium