Dev Biol 1987 Dec 01;4271:11-9.

GTP-binding proteins Gi and Go transplanted onto Xenopus oocyte by rat brain messenger RNA.

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After injection with messenger RNA (mRNA) isolated from rat brain, Xenopus laevis oocytes acquired electrophysiological responsiveness to externally perfused acetylcholine (ACh) or serotonin (5-HT), and elevated responsiveness to internally applied guanosine 5'-(3-O-thio)triphosphate (GTP gamma S). Compared with the membranes of native oocytes, those of mRNA-injected oocytes contained increased amounts of 39 and 41 kDa proteins, which could be [32P]ADP-ribosylated by pertussis toxin (PTX). The amplitude of the GTP gamma S response and the amounts of the 39 and 41 kDa proteins increased in a parallel manner for at least 3 days following mRNA injection. Current responses to internally applied GTP gamma S showed properties common to those of responses to ACh or 5-HT perfusion: both responses had reversal potentials close to the Cl- potential, were mimicked by intracellular injection of IP3, desensitized by a large dose of IP3, and inhibited by a simultaneous injection of neomycin or EGTA. Incubation of mRNA-injected cells with PTX inhibited both the 5-HT response and the [32P]ADP-ribosylation of the 39 and 41 kDa proteins in a parallel, dose-dependent manner. After pretreatment of oocytes with PTX followed by mRNA injection, the levels of the 39 and kDa proteins and the 5-HT response appeared to be similar to those of non-treated cells injected with mRNA, whereas no detectable amounts of these proteins were induced when PTX-pretreated cells were analyzed under the same conditions without mRNA injection.(ABSTRACT TRUNCATED AT 250 WORDS)

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