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3
Nicotinic Subunit Gene Promoter
CNR Cellular and Molecular Pharmacology Center, Department of
Medical Pharmacology, University of Milan, 20129 Milan, Italy
We describe the structural and functional features of the human 
3
nicotinic receptor subunit promoter. A 0.35-kb region immediately upstream of the start codon was identified that when transfected in
human neuroblastoma cells was able to drive the expression of the
luciferase reporter gene with a strength comparable to that of the
well-characterized simian virus 40 promoter/enhancer. This region
displayed the features of a multistart-site, GC-rich, TATA-less, and
CAAT-less promoter, containing many overlapping Sp1 and AP-2 putative
binding sites. Further dissections of the 0.35-kb fragment revealed
that its 3
region, specifying the 5 UT of the mRNA, plays a relevant
positive effect in determining the strength of the promoter. This
region contains putative cis-acting elements for AP-2,
nuclear factor-
B, and the recently described multiple-start site
element downstream-1. By mutation analysis, we showed that these sites
are functional and when combined increase the promoter activity by
4-fold. The 0.35-kb promoter was found to be under the negative control
of upstream sequences that include a modern Alu repeat. The 3 Alu
repeat works as a composite region, containing both positive and
negative elements that control the activity of the downstream promoter.
Finally, we investigated the tissue-specific activity of the human 3
gene 5 regulatory sequences, showing that they are able to drive the
expression of the reporter gene preferentially in neuronal cells.
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