Expression of the four subunits of the Torpedo californica nicotinic acetylcholine receptor in Saccharomyces cerevisiae

K. U. Jansen, W. G. Conroy, T. Claudio, T. D. Fox, N. Fujita, O. Hamill, J. M. Lindstrom, M. Luther, N. Nelson, K. A. Ryan, M. T. Sweet, G. P. Hess

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

Yeast expression vectors were constructed containing complementary DNA encoding the α-, β-, γ-, and δ-subunits of the Torpedo californica nicotinic acetylcholine receptor under the control of the Saccharomyces cerevisiae alcohol dehydrogenase promoter. All four plasmids were integrated into the yeast genome of a single yeast cell. The resulting yeast strain synthesized polypeptides novel to yeast that had the molecular weights and antigenic properties similar to the authentic T. californica receptor α-, γ-, and δ-subunits. The β-subunit polypeptide could not be detected in this yeast strain, even though the poly(A)+ RNA from this strain contained all the information necessary for the expression of functional acetylcholine receptors in Xenopus laevis oocytes. The replacement of the β-subunit mRNA 5'-untranslated leader and its N-terminal signal sequence by the corresponding α-subunit sequences, however, resulted in the expression of the β-subunit polypeptide in yeast grown at 5°C.

Original languageEnglish (US)
Pages (from-to)15022-15027
Number of pages6
JournalJournal of Biological Chemistry
Volume264
Issue number25
StatePublished - Jan 1 1989
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Jansen, K. U., Conroy, W. G., Claudio, T., Fox, T. D., Fujita, N., Hamill, O., Lindstrom, J. M., Luther, M., Nelson, N., Ryan, K. A., Sweet, M. T., & Hess, G. P. (1989). Expression of the four subunits of the Torpedo californica nicotinic acetylcholine receptor in Saccharomyces cerevisiae. Journal of Biological Chemistry, 264(25), 15022-15027.