Complementation in situ of the yeast plasma membrane H+‐ATPase gene pmal by an H+‐ATPase gene from a heterologous species

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Abstract
In plants and fungi, the transport of solutes across the plasma membrane (pm) is driven by a proton pump (H+-ATPase) that produces an electric potential and a pH gradient. We expressed AHA2, a member of the Arabidopsis thaliana pm H+-ATPase gene family, in yeast cells in which transcription of the endogenous pm H+ATPase gene (pmal) had been turned off. AHA2 was expressed mainly in intracellular membranes and only supported very slow growth of transformed yeast cells. Removal of the last 92 C-terminal amino acids from the plant H+-ATPase produced an enzyme with 2–3-fold higher specific ATPase activity than the wild-type plant enzyme. Surprisingly, the truncated H+-ATPase was now targetted to the yeast pm and fully supported normal yeast growth.