Molecular cloning and sequence determination of a cDNA coding for the α-subunit of a Go-type protein of Xenopus laevis oocytes

Abstract

Xenopus laevis oocytes are cells ideally suited to the study of signal transduction and of the G-proteins that are involved in this process. A X. laevis cDNA library in λgt10 has been screened with a mixture of three oligonucleotide probes designed to detect sequences found in various mammalian α-subunits of G-proteins. One of these clones has been purified through tertiary screening and the DNA insert has been sequenced. This clone was found to include the total sequence coding for a 354 amino acid protein that is 89% identical to the sequence of α-subunit of rat Go. The differences with the mammalian protein were clustered in amino acids 290-315, which have been postulated to define the region interacting with the receptor and effector molecule. The homology with the α-subunits of other mammalian G-proteins is lower (65-70% to Gi and 42% to Gs). On this basis, this clone can be classified as Go-like. © 1989.