Reference - Detail
|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|
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.
|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|
|WOS Category||GENETICS & HEREDITY BIOCHEMISTRY & MOLECULAR BIOLOGY|
|Yeast||KJ100-7B pREP41 pTN381(GFP-psy1)|