RRC ID 887
Author Iwaki T, Goa T, Tanaka N, Takegawa K.
Title Characterization of Schizosaccharomyces pombe mutants defective in vacuolar acidification and protein sorting.
Journal Mol. Genet. Genomics
Abstract The vacuolar H+-ATPases (V-ATPases) are ATP-dependent proton pumps responsible for acidification of intracellular compartments in eukaryotic cells. To investigate the functional roles of the V-ATPase in Schizosaccharomyces pombe, the gene vma1 encoding subunit A or vma3 encoding subunit c was disrupted. Both deletion mutants lost the capacity for vacuolar acidification in vivo, and showed sensitivity to neutral pH or high concentrations of divalent cations including Ca2+. The delivery of FM4-64 to the vacuolar membrane and accumulation of Lucifer Yellow CH were strongly inhibited in the vma1 and vma3 mutants. Moreover, deletion of the S. pombe vma1+ or vma3+ gene resulted in pleiotropic phenotypes consistent with lack of vacuolar acidification, including the missorting of vacuolar carboxypeptidase Y, abnormal vacuole morphology, and mating defects. These findings suggest that V-ATPase is essential for endocytosis, ion and pH homeostasis, and for intracellular targeting of vacuolar proteins and vacuolar biogenesis in S. pombe.
Volume 271(2)
Pages 197-207
Published 2004-3
DOI 10.1007/s00438-003-0971-7
PMID 14735354
MeSH Cathepsin A / metabolism DNA Primers Databases, Genetic Endocytosis / physiology Fluorescent Dyes / metabolism Gene Deletion Hydrogen-Ion Concentration Immunoblotting Isoquinolines / metabolism Microscopy, Fluorescence Mutation / genetics* Plasmids / genetics Protein Transport Pyridinium Compounds / metabolism Quaternary Ammonium Compounds / metabolism Schizosaccharomyces / enzymology Schizosaccharomyces / genetics* Sequence Homology, Amino Acid Vacuolar Proton-Translocating ATPases / genetics* Vacuolar Proton-Translocating ATPases / physiology* Vacuoles / ultrastructure
IF 2.879
Times Cited 26
WOS Category GENETICS & HEREDITY BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Yeast KJ100-7B pREP41 pTN381(GFP-psy1)