A single mutation confers vanadate resistance to the plasma membrane H+-ATPase from the yeast Schizosaccharomyces pombe.
A single-gene nuclear mutant has been selected from the yeast Schizosaccharomyces pombe for growth resistance to Dio-9, a plasma membrane H+-ATPase inhibitor. From this mutant, called pma1, an ATPase activity has been purified. It contains a Mr = 100,000 major polypeptide which is phosphorylated by [gamma-32P] ATP. Proton pumping is not impaired since the isolated mutant ATPase is able, in reconstituted proteoliposomes, to quench the fluorescence of the delta pH probe 9-amino-6-chloro-2-methoxy acridine. The isolated mutant ATPase is sensitive to Dio-9 as well as to seven other plasma membrane H+-ATPase inhibitors. The mutant H+-ATPase activity tested in vitro is, however, insensitive to vanadate. Its Km for MgATP is modified and its ATPase specific activity is decreased. The pma1 mutation decreases the rate of extracellular acidification induced by glucose when cells are incubated at pH 4.5 under nongrowing conditions. During growth, the intracellular mutant pH is more acid than the wild type one. The derepression by ammonia starvation of methionine transport is decreased in the mutant. The growth rate of pma1 mutants is reduced in minimal medium compared to rich medium, especially when combined to an auxotrophic mutation. It is concluded that the H+-ATPase activity from yeast plasma membranes controls the intracellular pH as well as the derepression of amino acid, purine, and pyrimidine uptakes. The pma1 mutation modifies several transport properties of the cells including those responsible for the uptake of Dio-9 and other inhibitors (Ulaszewski, S., Coddington, A., and Goffeau, A. (1986) Curr. Genet. 10, 359-364).
D000255:Adenosine Triphosphate / Q000378:metabolism; D001667:Binding, Competitive; D001692:Biological Transport; D002462:Cell Membrane / Q000201:enzymology; D004352:Drug Resistance, Microbial / Q000235:genetics; D005947:Glucose / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D009154:Mutation; D009930:Organic Chemicals; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D011522:Protons; D004718:Saccharomycetales / Q000235:genetics*; D012568:Schizosaccharomyces / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D014475:Uncoupling Agents / Q000494:pharmacology; D014638:Vanadates; D014639:Vanadium / Q000494:pharmacology*
D009930:Organic Chemicals; D011522:Protons; D014475:Uncoupling Agents; D014639:Vanadium; C100056:DIO 9; D014638:Vanadates; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases; D005947:Glucose