Basidiomycetous ras cDNA functionally replaces its homolog genes in yeast.

Abstract

It was shown by a plasmid exchange procedure that the Ras-encoding cDNA of the basidiomycete Lentinus edodes (named Leras cDNA) can functionally replace its homolog genes (ScRAS1 and ScRAS2) in the yeast Saccharomyces cerevisiae to maintain the viability of an yeast strain containing genetic disruptions of both RAS genes. The strain replaced by a Leras-cDNA-carrying plasmid, however, grew slower than the strains replaced by a ScRAS1- or a ScRAS2-carrying plasmid. The intracellular level of cAMP in the strain harboring the Leras-cDNA-carrying plasmid was clearly higher than that of a parental strain which maintains a plasmid carrying the S. cerevisiae cAMP-dependent protein kinase catalytic subunit C1 gene, TPK1, but was lower than that in a strain harboring an ScRAS2-carrying plasmid. These results suggest that the Leras cDNA can complement the ras1- ras2- mutation of yeast by virture of the stimulation of adenylate cyclase activity, although the complementation is not as efficient as that obtained by expressing the ScRAS2 gene.