Cell de-energization prevents plasmid transformation of yeast Saccharomyces cerevisiae: evidence for the requirement of ATP

Abstract

AbstractThe dependence of the yeast Saccharomyces cerevisiae transformation on energy requirement was studied. The inhibitory effect of sodium arsenate, used for the depletion of the intracellular ATP pool, was determined. Incubation of the yeast cells in 5 mM sodium arsenate diminished ATP accumulation by 50% and the transformation efficiency decreased by 65%. To discriminate between ATP produced by substrate level phosphorylation and oxidative phosphorylation, the inhibitory analysis of a mutant with defective mitochondria was performed. Sodium fluoride (10–50 mM), as inhibitor of glycolysis, elicited a concentration-dependent decrease in intracellular ATP levels in both parental and mutant cells. The equal transformation efficiency of the mitochondrial mutant and parental strain, in addition to experiments with oligomycin, demonstrated the independence of plasmid transformation on mitochondrial ATP synthesis. This is consistent with our hypothesis that yeast transformation efficiency is associated with ATP produced by substrate level phosphorylation.