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J. Biol. Chem., Vol. 265, Issue 8, 4472-4482, Mar, 1990
J Boulter, A O'Shea-Greenfield, RM Duvoisin, JG Connolly, E Wada, A Jensen, PD Gardner, M Ballivet, ES Deneris and D McKinnon
We have identified two additional members of the neuronal nicotinic
acetylcholine receptor (nAChR)-related gene family. cDNA clones for one new
gene, designated alpha 5, were isolated from rat hippocampus and rat PC12
cell line cDNA libraries. The alpha 5 gene encodes a protein of 48,800
daltons (424 amino acids) which exhibits significant overall amino acid
sequence identity with the previously cloned rat nAChR subunits alpha 1
(49%), alpha 2 (55%), alpha 3 (52%), and alpha 4 (49%). Features
characteristic of other nAChR alpha-subunits are present such as conserved
cysteine residues at positions 127, 141, 191, and 192, and four strongly
hydrophobic domains. A second addition to the nAChR-related gene family,
designated beta 4, is encoded in overlapping rat genomic clones lambda DD15
and lambda RG518A. The beta 4 gene, encoding a mature protein of 53,300
daltons (475 amino acids), consists of 6 exons and has a transcription unit
length of approximately 18 kilobase pairs. The beta 4 gene encoded protein
shows considerable amino acid sequence identity with nAChR beta 1 (43%),
beta 2 (64%), and beta 3 (44%) subunits. Northern blots showed that, along
with alpha 3 and beta 2, transcripts for both the alpha 5 and beta 4 genes
are present in the PC12 cell line, while in situ hybridization experiments
demonstrated expression of the alpha 5 and beta 4 genes in a small number
of nuclei in the central nervous system. Finally, the genes that encode the
beta 4, alpha 3, and alpha 5 proteins are transcribed with convergent
polarities and form a tightly linked gene cluster spanning approximately 60
kilobase pairs.
Alpha 3, alpha 5, and beta 4: three members of the rat neuronal nicotinic acetylcholine receptor-related gene family form a gene cluster
Molecular Neurobiology Laboratory, Salk Institute, San Diego, California 92138.
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