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J. Biol. Chem., Vol. 276, Issue 20, 16749-16757, May 18, 2001
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7 Promoter*
§,, and
From the Departments of ¶ Neuroscience and
Pharmacology, Neuronal nicotinic acetylcholine receptors
(nAChRs) containing the The Ohio State University College of
Medicine and Public Health, Columbus, Ohio 43210
7 subunit are expressed in the central
nervous system, autonomic nervous system, retina, adrenal medulla, and
PC12 cells. 7 nAChRs have been implicated in several important
biological activities apart from synaptic transmission such as
mediating neurite growth and presynaptic control of neurotransmitter
release. A 178-base pair promoter was sufficient to drive high level
expression of the 7 gene in PC12 cells. The 7 promoter was also
cell-specific, expressing in PC12 cells but not in L6 rat muscle cells.
Within our minimal rat 7 nAChR promoter we identified two sequences important for basal level expression. Mutation of a GC-rich sequence at
172 relative to the translational start site led to an increase in
activity of the promoter, indicating the presence of a negative regulatory element. Upstream stimulatory factor-1 acted to regulate 7 expression positively by binding to an E-box at 116. A site directly adjacent to the upstream stimulatory factor-1 binding site was
shown to bind Egr-1. Sp1 and Sp3 binding also occurred downstream from
or overlapping the Egr-1 binding site in the rat 7 promoter. Several
transcription factors interact in close proximity to control expression
of the rat 7 nicotinic receptor gene.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF321242.
§ Present address: Dept. of Dermatology, 1201 Welch Rd., MSLS Bldg., Stanford University, Stanford, CA 94305.
To whom correspondence should be addressed: Dept. of
Neuroscience, 4068 Graves Hall, Ohio State University College of
Medicine and Public Health, 333 West Tenth Ave., Columbus, OH
43210. Tel.: 614-292-4391; Fax: 614-688-8742; E-mail:
boyd.16@osu.edu.
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