Maturation of the yeast plasma membrane [H+]ATPase involves phosphorylation during intracellular transport.
In this study we show that the plasma membrane [H+]ATPase of Saccharomyces cerevisiae is phosphorylated on multiple Ser and Thr residues in vivo. Phosphorylation occurs during the movement of newly synthesized ATPase from the ER to the cell surface, as revealed by the analysis of temperature-sensitive sec mutants blocked at successive steps of the secretory pathway. Two-dimensional phosphopeptide analysis of the ATPase indicates that, although most sites are phosphorylated at or before arrival in secretory vesicles, some phosphopeptides are unique to the plasma membrane. Phosphorylation of plasma membrane-specific site(s) is associated with increased ATPase activity during growth on glucose. Upon glucose starvation, dephosphorylation occurs concomitantly with a decrease in enzymatic activity, and both are rapidly reversed (within 2 min) upon readdition of glucose. We suggest that reversible, site-specific phosphorylation serves to adjust ATPase activity in response to nutritional signals.
Chang
A
A
Department of Human Genetics, Yale University School of Medicine, New Haven, Connecticut 06510.
Slayman
C W
CW
D001692:Biological Transport / Q000502:physiology; D015153:Blotting, Western; D004721:Endoplasmic Reticulum / Q000378:metabolism; D004789:Enzyme Activation / Q000502:physiology*; D005656:Fungal Proteins / Q000378:metabolism*; D005947:Glucose / Q000494:pharmacology; D007700:Kinetics; D008565:Membrane Proteins / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D006180:Proton-Translocating ATPases / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D012694:Serine / Q000378:metabolism; D013696:Temperature; D013912:Threonine / Q000378:metabolism
D005656:Fungal Proteins; D008565:Membrane Proteins; D010750:Phosphoproteins; D013912:Threonine; D012694:Serine; D006180:Proton-Translocating ATPases; D005947:Glucose