Diploids heterozygous for a vma13Delta mutation in Saccharomyces cerevisiae highlight the importance of V-ATPase subunit balance in supporting vacuolar acidification and silencing cytosolic V1-ATPase activity.
The V-ATPase H subunit (encoded by the VMA13 gene) activates ATP-driven proton pumping in intact V-ATPase complexes and inhibits MgATPase activity in cytosolic V1 sectors (Parra, K. J., Keenan, K. L., and Kane, P. M. (2000) J. Biol. Chem. 275, 21761-21767). Yeast diploids heterozygous for a vma13Delta mutation show the pH- and calcium-dependent conditional lethality characteristic of mutants lacking V-ATPase activity, although they still contain one wild-type copy of VMA13. Vacuolar vesicles from this strain have approximately 50% of the ATPase activity of those from a wild-type diploid but do not support formation of a proton gradient. Compound heterozygotes with a second heterozygous deletion in another V1 subunit gene exhibit improved growth, vacuolar acidification, and ATP-driven proton transport in vacuolar vesicles. In contrast, compound heterozygotes with a second deletion in a Vo subunit grow even more poorly than the vma13Delta heterozygote, have very little vacuolar acidification, and have very low levels of V-ATPase subunits in isolated vacuoles. In addition, cytosolic V1 sectors from this strain and from the strain containing only the heterozygous vma13Delta mutation have elevated MgATPase activity. The results suggest that balancing levels of subunit H with those of other V-ATPase subunits is critical, both for allowing organelle acidification and for preventing unproductive hydrolysis of cytosolic ATP.
Rizzo
Jason M
JM
Department of Biochemistry and Molecular Biology, State University of New York Upstate Medical University, Syracuse, New York 13210, USA.
Tarsio
Maureen
M
Martínez-Muñoz
Gloria A
GA
Kane
Patricia M
PM
D000251:Adenosine Triphosphatases / Q000378:metabolism; D017301:Ca(2+) Mg(2+)-ATPase / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology* / Q000378:metabolism; D004171:Diploidy; D017353:Gene Deletion; D020868:Gene Silencing; D005838:Genotype; D006579:Heterozygote; D006868:Hydrolysis; D018942:Macrolides / Q000494:pharmacology; D008954:Models, Biological; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics* / Q000502:physiology; D025262:Vacuolar Proton-Translocating ATPases / Q000737:chemistry* / Q000235:genetics* / Q000502:physiology
D018942:Macrolides; D029701:Saccharomyces cerevisiae Proteins; C447617:VMA13 protein, S cerevisiae; C036978:concanamycin A; D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase; D025262:Vacuolar Proton-Translocating ATPases