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PMID: 2141385 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Role of vacuolar acidification in protein sorting and zymogen activation: a genetic analysis of the yeast vacuolar proton-translocating ATPase.

Molecular and cellular biology ·Vol. 10 ·No. 7 ·1990-07-00 ·Pages 3737-49

Yamashiro CT, Kane PM, Wolczyk DF, Preston RA, Stevens TH

Abstract

Vacuolar acidification has been proposed to play a key role in a number of cellular processes, including protein sorting, zymogen activation, and maintenance of intracellular pH. We investigated the significance of vacuolar acidification by cloning and mutagenizing the gene for the yeast vacuolar proton-translocating ATPase 60-kilodalton subunit (VAT2). Cells carrying a vat2 null allele were viable; however, these cells were severely defective for growth in medium buffered at neutral pH. Vacuoles isolated from cells bearing the vat2 null allele were completely devoid of vacuolar ATPase activity. The pH of the vacuolar lumen of cells bearing the vat2 mutation was 7.1, compared with the wild-type pH of 6.1, as determined by a flow cytometric pH assay. These results indicate that the vacuolar proton-translocating ATPase complex is essential for vacuolar acidification and that the low-pH state of the vacuole is crucial for normal growth. The vacuolar acidification-defective vat2 mutant exhibited normal zymogen activation but displayed a minor defect in vacuolar protein sorting.

MeSH Terms
Amino Acid Sequence Base Sequence Cloning, Molecular Enzyme Activation Enzyme Precursors/genetics Escherichia coli/genetics Genes, Fungal Genotype Hydrogen-Ion Concentration Macromolecular Substances Molecular Sequence Data Mutation Oligonucleotide Probes Proton-Translocating ATPases/genetics Restriction Mapping Saccharomyces cerevisiae/enzymology,genetics Sequence Homology, Nucleic Acid Vacuoles/enzymology
Chemicals
Enzyme Precursors Macromolecular Substances Oligonucleotide Probes Proton-Translocating ATPases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Yamashiro C T
Institute of Molecular Biology, University of Oregon, Eugene 97403.
Kane P M
Wolczyk D F
Preston R A
Stevens T H
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.10.7.3737-3749.1990
Published
1990-07-00
Pages
3737-49
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC360825
Subset
IM
Grants
NIGMS NIH HHS · GM11329 · United States
NIGMS NIH HHS · GM29713 · United States
NIGMS NIH HHS · GM32508 · United States
External Links
PubMed source DOI: 10.1128/mcb.10.7.3737-3749.1990 PMC Full Text View journal details
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