Please use this identifier to cite or link to this item: doi:10.22028/D291-40648
Title: Remodeling of yeast vacuole membrane lipidomes from the log (one phase) to stationary stage (two phases)
Author(s): Reinhard, John
Leveille, Chantelle L.
Cornell, Caitlin E.
Merz, Alexey J.
Klose, Christian
Ernst, Robert
Keller, Sarah L.
Language: English
Title: Biophysical Journal
Volume: 122
Issue: 6
Pages: 1043-1057
Publisher/Platform: Elsevier
Year of Publication: 2023
DDC notations: 610 Medicine and health
Publikation type: Journal Article
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.
DOI of the first publication: 10.1016/j.bpj.2023.01.009
URL of the first publication:
Link to this record: urn:nbn:de:bsz:291--ds-406480
ISSN: 0006-3495
Date of registration: 28-Sep-2023
Description of the related object: Supporting material
Related object:
Faculty: M - Medizinische Fakultät
Department: M - Medizinische Biochemie und Molekularbiologie
Professorship: M - Prof. Dr. Robert Ernst
Collections:SciDok - Der Wissenschaftsserver der Universität des Saarlandes

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