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Volume 17, Number 7,
Issue of April 1, 1997
pp. 2273-2283
Copyright ©1997 Society for Neuroscience
 43 : An Enhancer Displaying Neural-Restricted Activity Is
Located in the 3-Untranslated Exon of the Rat Nicotinic Acetylcholine
Receptor 4 Gene
Received Aug. 20, 1996; revised Jan. 14, 1997; accepted Jan. 15, 1997.
Jennifer McDonough and
Evan Deneris
Department of Neurosciences, School of Medicine, Case Western
Reserve University, Cleveland, Ohio 44106
Members of a neuronal nicotinic acetylcholine receptor subunit gene
cluster ordered  4,  3, 5 in the vertebrate genome are expressed
in highly restricted patterns in the PNS and CNS. Nothing is known,
however, about the regulatory elements that control transcription of
these genes in selected neuronal cell populations. We report here a
novel enhancer, designated 43 , that is positioned in the 4
3-untranslated exon. It is composed of two nearly identical 37 bp
direct repeats that are separated by 6 bp. Multimerization of the
enhancer upstream of the 3 minimal promoter results in synergistic
activation. Analysis in different cell types, including three neural
lines and primary keratinocytes, shows that 43 is preferentially
active in the neural line PC12, which expresses all members of the
cluster. Mobility shift assays reveal a cell-type-specific complex,
which forms with the first repeat of the enhancer and PC12 extracts.
Complexes co-migrating with the PC12 cell complex are not detected with
extracts from other lines, which suggests that PC12 cells contain a
differentially expressed factor that may be important for the
restricted activity of 43. The cell-type-specific activity of the
43 enhancer suggests that it is important for regulating restricted
expression patterns of one or more clustered neuronal acetylcholine
receptor genes. Its location within the 4 gene may be a selective
pressure for maintaining tight linkage of clustered neuronal nAchR
genes.
Key words:
neuronal nicotinic receptors;
cis-acting elements;
enhancer;
cell-type specific;
gene transcription;
3-untranslated
exon
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