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Control of Torpedo Acetylcholine Receptor Biosynthesis in Xenopus Oocytes
Journal article   Open access   Peer reviewed

Control of Torpedo Acetylcholine Receptor Biosynthesis in Xenopus Oocytes

Amy L. Buller and Michael M. White
Proceedings of the National Academy of Sciences - PNAS, v 85(22), pp 8717-8721
01 Nov 1988
PMID: 3186754
url
https://doi.org/10.1073/pnas.85.22.8717View
Published, Version of Record (VoR) Open

Abstract

Biosynthesis Cell surface receptors Complementary DNA Microinjections Oocytes Plasmids Receptors RNA Stoichiometry Teeth
The nicotinic acetylcholine receptor (AcChoR) of Torpedo electroplax is a multisubunit transmembrane glycoprotein complex with a subunit stoichiometry of α 2β γ δ . The RNAs for the separate subunits were transcribed in vitro from cDNAs inserted in pSP64T vectors and microinjected in Xenopus oocytes. Microinjected in vitro-transcribed RNAs were stable, with a half-life of 72 hr. Xenopus oocytes assembled functional AcChoRs from the subunit-specific RNAs. These receptors were inserted in the cell membrane and could be detected as early as 6 hr after RNA microinjection. The oocyte-expressed AcChoR subunits could be immunoprecipitated with anti-Torpedo AcChoR subunit antibodies. Expression of the AcChoR in oocytes proceeded linearly for 72 hr after microinjection. While the amount of RNA injected did not alter the linearity of the expression time course, the rate of receptor expression in oocytes showed a saturable dependence on RNA concentration. Varying the relative amount of α -subunit RNA microinjected into oocytes had a striking effect on receptor expression. This effect was specific for the α -subunit. These results suggest that transcript availability may control receptor expression in Xenopus oocytes. In addition, the availability of the α -subunit may be a limiting factor for receptor expression.

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Biochemistry & Molecular Biology
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