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Published April 25, 1986 | public
Journal Article Open

Mouse muscle nicotinic acetylcholine receptor gamma subunit: cDNA sequence and gene expression

Abstract

Clones coding for the mouse nicotinic acetylcholine receptor (AChR) gamma subunit precursor have been selected from a cDNA library derived from a mouse myogenic cell line and sequenced. The deduced protein sequence consists of a signal peptide of 22 amino acid residues and a mature gamma subunit of 497 amino acid residues. There is a high degree of sequence conservation between this mouse sequence and published human and calf AChR gamma subunits and, after allowing for functional amino acid substitutions, also to the more distantly related chicken and Torpedo AChR gamma subunits. The degree of sequence conservation is especially high in the four putative hydrophobic membrane spanning regions, supporting the assignment of these domains. RNA blot hybridization showed that the mRNA level of the gamma subunit increases by 30 fold or more upon differentiation of the two mouse myogenic cell lines, BC3H-1 and C2C12, suggesting that the primary controls for changes in gene expression during differentiation are at the level of transcription. One cDNA clone was found to correspond to a partially processed nuclear transcript containing two as yet unspliced intervening sequences.

Additional Information

© 1986 IRL Press Limited, Oxford, England. Received 19 December 1985; Accepted 6 February 1986. We are grateful to Ms. Marie Krempin for operation of the cell culture facility, to Dr. Terry Snutch for providing the RNA purification procedure, to Tim Hunkapiller for the usage of computer sequence analysis system (48), and to Dr. Jean-Paul Revel for the usage of graphic digitizer. The Torpedo γ probe was kindly provided by the authors of reference 8. While this manuscript was in the final stages of preparation, we received a copy of a preprint from Dr. J. Boulter and coworkers in the laboratory of Drs. S. Heinemann and J. Patrick at the Salk Institute describing their independent isolation and sequence determination of a γ subunit cDNA clone derived from BC3H-1 cells. There are only a few minor disagreements between the two sequences. We are grateful to these authors for sharing their results prior to publication. This work was supported by research grants to N. D. from the National Institutes of Health and the Muscular Dystrophy Association, by a California Foundation for Biochemical Research Fellowship to L.Y., and by a National Institutes of Health Fellowship to R.L.P.

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August 22, 2023
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October 16, 2023