Cloning and characterization of a cDNA coding for the alpha-subunit of a stimulatory G protein from Schistosoma mansoni.
Guanine nucleotide-binding proteins (G proteins) mediate signals between serotonin receptors and adenylate cyclase in Schistosoma mansoni. A bovine Gs alpha cDNA probe was used to isolate a cDNA clone, SG12, encoding the entire alpha-subunit of a G protein of S. mansoni. The cDNA is 1897 base pairs long, contains an open reading frame of 1137 base pairs, and codes for a deduced protein of 379 amino acids. The putative protein encoded by the clone has an exact amino acid match with bovine Gs alpha of 65% and a 78% match when conserved amino acid substitutions are considered. In contrast, the exact and conserved matches of the schistosome alpha-subunit with bovine Gi are 41 and 61%, respectively. A comparison of the deduced amino acid sequence of SG12 with a variety of different G alpha proteins indicates that all the major structural features characteristic of a Gs alpha protein are present in the S. mansoni gene. The schistosome clone contains the putative site for ADP-ribosylation by cholera toxin found in Gs alpha but does not contain the ADP-ribosylation site for pertussis toxin present in Gi alpha. The amino acids are completely conserved at the GTP-binding sites. On a Northern blot, the cDNA hybridizes to a major band of 3.1 kilobases in RNA from adult schistosomes. The message appears to be absent in miracidia and cercariae, but a faint 3.1-kilobase band is visible in the early schistosomule stage preceding adulthood. This evidence, when added to previous biochemical data, indicates that the expression of this gene is developmentally controlled.