Characterization of a fission yeast gene, gpa2, that encodes a G alpha subunit involved in the monitoring of nutrition

Abstract

The Schizosaccharomyces pombe gpa2 gene was cloned by hybridization with a cDNA for Dictyostelium discoideum G alpha 1. It encodes a homolog of G-protein alpha-subunits with 354 amino acids and a predicted molecular mass of 40,522. Disruption of gpa2 slows cell growth but is not lethal. Cells defective in gpa2 mate and sporulate readily in the presence of plentiful nutrition, bypassing the requirement of nitrogen starvation for the initiation of sexual development. These phenotypes mimic those of cells defective in cyr1 encoding adenylyl cyclase. The level of cAMP in gpa2 null mutants is only one-third of the wild-type level. Mutations in gpa2 that are likely to inhibit the GTPase activity of the gene product cause a slight increase in intracellular cAMP levels and result in leaky sterility. The cAMP level reaches 20 times as high as the wild-type level if a cell carries both this type of gpa2 mutation and a null mutation in pde1 encoding phosphodiesterase. Cells defective in gpa2 fail to produce cAMP in response to glucose stimulation. These results suggest that Gpa2 is involved in the determination of the cAMP level according to nutritional conditions, most likely as a positive regulator of adenylyl cyclase.