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• W4315652371 endingPage "1057" @default.
• W4315652371 startingPage "1043" @default.
• W4315652371 abstract "Upon nutrient limitation, budding yeast of Saccharomyces cerevisiae shift from fast growth (the log stage) to quiescence (the stationary stage). This shift is accompanied by liquid-liquid phase separation in the membrane of the vacuole, an endosomal organelle. Recent work indicates that the resulting micrometer-scale domains in vacuole membranes enable yeast to survive periods of stress. An outstanding question is which molecular changes might cause this membrane phase separation. Here, we conduct lipidomics of vacuole membranes in both the log and stationary stages. Isolation of pure vacuole membranes is challenging in the stationary stage, when lipid droplets are in close contact with vacuoles. Immuno-isolation has previously been shown to successfully purify log-stage vacuole membranes with high organelle specificity, but it was not previously possible to immuno-isolate stationary-stage vacuole membranes. Here, we develop Mam3 as a bait protein for vacuole immuno-isolation, and demonstrate low contamination by non-vacuolar membranes. We find that stationary-stage vacuole membranes contain surprisingly high fractions of phosphatidylcholine lipids (∼40%), roughly twice as much as log-stage membranes. Moreover, in the stationary stage, these lipids have higher melting temperatures, due to longer and more saturated acyl chains. Another surprise is that no significant change in sterol content is observed. These lipidomic changes, which are largely reflected on the whole-cell level, fit within the predominant view that phase separation in membranes requires at least three types of molecules to be present: lipids with high melting temperatures, lipids with low melting temperatures, and sterols." @default.
• W4315652371 created "2023-01-12" @default.
• W4315652371 creator A5000338874 @default.
• W4315652371 creator A5021355660 @default.
• W4315652371 creator A5027385934 @default.
• W4315652371 creator A5027410618 @default.
• W4315652371 creator A5042037721 @default.
• W4315652371 creator A5062782296 @default.
• W4315652371 creator A5071326290 @default.
• W4315652371 date "2023-03-01" @default.
• W4315652371 modified "2023-10-09" @default.
• W4315652371 title "Remodeling of yeast vacuole membrane lipidomes from the log (one phase) to stationary stage (two phases)" @default.
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