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Volume 272, Number 8, Issue of February 21, 1997 pp. 5098-5104
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

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Isolation and Characterization of the 5-Upstream Region of the Human N-type Calcium Channel _1B Subunit Gene
CHROMOSOMAL LOCALIZATION AND PROMOTER ANALYSIS

(Received for publication, June 25, 1996, and in revised form, October 30, 1996)

Dong S. Kim , Hyun-Ho Jung , Sun-Hwa Park ^§ and Hemin Chin

From the  Laboratory of Neurochemistry, NINDS, National Institutes of Health, Bethesda, Maryland 20892 and ^§ Department of Anatomy, College of Medicine, Korea University, Seoul, Korea 136-701

-Conotoxin-sensitive N-type Ca²⁺ channels, unlike dihydropyridine-sensitive L-type channels, are exclusively expressed in nervous tissues. To understand the molecular basis for neuron-specific expression of the N-type channel, we have isolated genomic clones encoding the human _1B subunit gene, localized to the long arm of chromosome 9 (9q34) by fluorescence in situ hybridization, and characterized its 5-upstream region. The proximal promoter of the _1B subunit gene lacks a typical TATA box, is highly GC-rich, and contains several sequences for transcription factor binding. Primer extension experiments revealed the presence of two transcription start sites. In vitro transfection study of the _1B subunit-luciferase fusion gene showed that the 4.0-kb 5-flanking region of the _1B gene functions as an efficient promoter in neuronal cells but not in glioma or nonneuronal cells, consistent with the patterns of the endogenous _1B gene expression in these cells. Deletion analysis of _1B subunit-luciferase fusion gene constructs further revealed the presence of several cis-acting regulatory elements, including a potential repressor located in the distal upstream region (3992 to 1788) that may be important for the neuron-specific expression of the N-type Ca²⁺ channel _1B subunit gene.

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