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Complementation in situ of the yeast plasma membrane H + ‐ATPase gene pmal by an H + ‐ATPase gene from a heterologous species
Author(s) -
Palmgren Michael Gjedde,
Christensen Gertrud
Publication year - 1993
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(93)81279-9
Subject(s) - yeast , complementation , atpase , biochemistry , heterologous expression , enzyme , biology , saccharomyces cerevisiae , arabidopsis thaliana , heterologous , gene , membrane , microbiology and biotechnology , chemistry , mutant , recombinant dna
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.