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Measurement of Vacuolar and Cytosolic pH In Vivo in Yeast Cell Suspensions
Published on: April 19, 2013
Nystatin effects on vacuolar function in Saccharomyces cerevisiae
M S Bhuiyan1, Y Ito, A Nakamura
1Department of Life Sciences, Faculty of Agriculture, Kagawa University, Japan.
Bioscience, Biotechnology, and Biochemistry
|July 31, 1999
Summary
Nystatin antibiotic sensitivity in yeast (Saccharomyces cerevisiae) reveals critical vacuolar sorting and morphology defects. Mutations in VPS16 gene cause nystatin sensitivity, highlighting its role in vacuolar function.
Area of Science:
- * Cell Biology
- * Microbiology
- * Genetics
Background:
- * Nystatin is a polyene antibiotic impacting fungal cell membranes.
- * Vacuolar protein sorting is crucial for cellular homeostasis in yeast (Saccharomyces cerevisiae).
- * Understanding nystatin's cellular effects can elucidate vacuolar functions.
Purpose of the Study:
- * To investigate the effects of nystatin on Saccharomyces cerevisiae.
- * To isolate and characterize nystatin-sensitive mutants to understand vacuolar protein sorting.
- * To identify the genetic basis of nystatin sensitivity in yeast.
Main Methods:
- * Ethylmethane sulfonate mutagenesis was used to generate nystatin-sensitive mutants.
- * Characterization of a specific mutant (nss1) under various stress conditions.
- * Analysis of vacuolar protein sorting and morphology in wild-type and mutant yeast strains.
- * Genetic analysis to identify the mutated gene in nystatin-sensitive strains.
Main Results:
- * Isolated and characterized nystatin-sensitive mutants, including the nss1 mutant.
- * The nss1 mutant exhibited sensitivity to heat and high cation concentrations.
- * Severe defects in vacuolar protein sorting and morphology were observed in nss1 mutants.
- * The nss1 mutation was mapped to the VPS16 gene, essential for vacuolar protein sorting.
- * All tested vacuolar deficient mutants (vps11, vps16, vps18, vps33) were sensitive to nystatin.
- * Nystatin treatment induced significant vacuole enlargement in wild-type yeast.
Conclusions:
- * Nystatin sensitivity in Saccharomyces cerevisiae is linked to defects in vacuolar protein sorting and morphology.
- * The VPS16 gene plays a critical role in maintaining vacuolar integrity and function.
- * Nystatin can be a valuable tool for studying vacuolar biology and identifying related genetic components.
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