Complementation in situ of the yeast plasma membrane H(+)-ATPase gene pma1 by an H(+)-ATPase gene from a heterologous species.
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 (pma1) 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.
Palmgren
M G
MG
August Krogh Institute, University of Copenhagen, Denmark.
Christensen
G
G
D017360:Arabidopsis / Q000235:genetics; D001483:Base Sequence; D002462:Cell Membrane / Q000201:enzymology; D004789:Enzyme Activation; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D006180:Proton-Translocating ATPases / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D013347:Subcellular Fractions / Q000201:enzymology; D014170:Transformation, Genetic
D006180:Proton-Translocating ATPases