Cell de-energization prevents plasmid transformation of yeast <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>: evidence for the requirement of ATP |
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Authors: | Larisa Chaustova Valė Miliukienė Aurelijus Zimkus |
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Institution: | (1) Key Laboratory of MOE for Plant Developmental Biology, College of Life Sciences, Wuhan University, Wuhan, Hubei, 430072, China; |
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Abstract: | The 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. |
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