Structural Role of Plasma Membrane Sterols in Osmotic Stress Tolerance of Yeast Saccharomyces cerevisiae
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals
2.2. Large Unilamellar Vesicle Preparation
2.3. Stopped Flow Experiments
2.4. GUV Experiments
2.5. Yeast Strains and Growth Conditions
2.6. Deletion of LAM2
2.7. Growth Kinetics Analysis
2.8. Propidium Iodide Uptake upon Hypoosmotic Stress
3. Results
3.1. Lowering Sterol Content of Artificial Liposomes Increases Their Water Permeability
3.2. Sterol Prevents Shrinkage of Giant Unilamellar Vesicles upon Transient Hyperosmotic Stress
3.3. Yeast Mutants in Sterol Biosynthesis/Trafficking Are More Sensitive to Sodium Chloride than to Sorbitol
3.4. Yeast Cells Deficient in the Reverse Transport of Sterol from the PM Accumulate Propidium Iodide upon Hypoosmotic Stress
4. Discussion
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
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Strain | Genotype | Parental Strain | Reference |
---|---|---|---|
W303 | MATa ade2–101 his3–11 trp1–1 ura3–52 can1–100 leu2–3 | W303 | Laboratory of A. Hyman |
UPC2-1 | MATa UPC2–1 ura3–1 his3–11,- 15 leu2–3,-112 trp1–1 | W303 | [23] |
UPC2-1Δlam2 | MATa UPC2–1 ura3–1 his3–11,- 15 leu2–3,-112 trp1–1Δlam2::TRP1 | W303 | This study |
Δlam1Δlam2Δlam3Δlam4 | MATa ade2–101 his3–11 trp1–1 ura3–52 can1–100 leu2–3 MATa ade2–101 his3–11 trp1–1 ura3– 52 can1–100 leu2–3Δlam3::kanMX4Δlam2::TRP1 | W303 | [23] |
BY4742 | MATalpha his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 | BY4742 | Deletion collection [24] |
Δhog1 | MATalpha his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 hog1::kanMX4 | BY4742 | Deletion collection [24] |
Δerg4Δlam2 | MATalpha his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 erg4::kanMX4Δlam2::HIS3 | BY4742 | [25] |
GUV Composition | 15 s from the Start of Application of the Hyperosmotic Solution | 30 s from the Start of Application of the Hyperosmotic Solution |
---|---|---|
DOPC | 12 ± 5% | 3 ± 2% |
70 mol.% DOPC + 30 mol./% Ergosterol | 5 ± 3% | 2 ± 1% |
70 mol.% DOPC + 30 mol./% Cholesterol | 4 ± 2% | 3 ± 2% |
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Sokolov, S.S.; Popova, M.M.; Pohl, P.; Horner, A.; Akimov, S.A.; Kireeva, N.A.; Knorre, D.A.; Batishchev, O.V.; Severin, F.F. Structural Role of Plasma Membrane Sterols in Osmotic Stress Tolerance of Yeast Saccharomyces cerevisiae. Membranes 2022, 12, 1278. https://doi.org/10.3390/membranes12121278
Sokolov SS, Popova MM, Pohl P, Horner A, Akimov SA, Kireeva NA, Knorre DA, Batishchev OV, Severin FF. Structural Role of Plasma Membrane Sterols in Osmotic Stress Tolerance of Yeast Saccharomyces cerevisiae. Membranes. 2022; 12(12):1278. https://doi.org/10.3390/membranes12121278
Chicago/Turabian StyleSokolov, Svyatoslav S., Marina M. Popova, Peter Pohl, Andreas Horner, Sergey A. Akimov, Natalia A. Kireeva, Dmitry A. Knorre, Oleg V. Batishchev, and Fedor F. Severin. 2022. "Structural Role of Plasma Membrane Sterols in Osmotic Stress Tolerance of Yeast Saccharomyces cerevisiae" Membranes 12, no. 12: 1278. https://doi.org/10.3390/membranes12121278