Regulation of vacuolar proton-translocating ATPase activity and assembly by extracellular pH.
Vacuolar proton-translocating ATPases (V-ATPases) are responsible for organelle acidification in all eukaryotic cells. The yeast V-ATPase, known to be regulated by reversible disassembly in response to glucose deprivation, was recently reported to be regulated by extracellular pH as well (Padilla-López, S., and Pearce, D. A. (2006) J. Biol. Chem. 281, 10273-10280). Consistent with those results, we find 57% higher V-ATPase activity in vacuoles isolated after cell growth at extracellular pH of 7 than after growth at pH 5 in minimal medium. Remarkably, under these conditions, the V-ATPase also becomes largely insensitive to reversible disassembly, maintaining a low vacuolar pH and high levels of V(1) subunit assembly, ATPase activity, and proton pumping during glucose deprivation. Cytosolic pH is constant under these conditions, indicating that the lack of reversible disassembly is not a response to altered cytosolic pH. We propose that when alternative mechanisms of vacuolar acidification are not available, maintaining V-ATPase activity becomes a priority, and the pump is not down-regulated in response to energy limitation. These results also suggest that integrated pH and metabolic inputs determine the final assembly state and activity of the V-ATPase.
Diakov
Theodore T
TT
Department of Biochemistry and Molecular Biology, SUNY Upstate Medical University, Syracuse, NY 13219, USA.
Kane
Patricia M
PM
D002462:Cell Membrane / Q000201:enzymology; D003600:Cytosol / Q000201:enzymology / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods; D005658:Fungi / Q000201:enzymology*; D015971:Gene Expression Regulation, Enzymologic; D005947:Glucose / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008954:Models, Biological; D009154:Mutation; D011522:Protons; D013997:Time Factors; D025262:Vacuolar Proton-Translocating ATPases / Q000737:chemistry*; D014617:Vacuoles / Q000201:enzymology
D011522:Protons; D025262:Vacuolar Proton-Translocating ATPases; D005947:Glucose