Open AccessVacuole-Specific Lipid Release for Tracking Intracellular Lipid Metabolism and Transport in Saccharomyces cerevisiae
Author(s) -
Vladimir Girik,
Suihan Feng,
Hanaa Hariri,
W. Mike Henne,
Howard Riezman
Publication year - 2022
Publication title -
acs chemical biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.899
H-Index - 111
eISSN - 1554-8937
pISSN - 1554-8929
DOI - 10.1021/acschembio.2c00120
Subject(s) - vacuole , saccharomyces cerevisiae , sphingolipid , endoplasmic reticulum , lipid metabolism , biochemistry , lipid droplet , microbiology and biotechnology , intracellular , ergosterol , yeast , biology , metabolism , lipotoxicity , chemistry , cytoplasm , insulin resistance , insulin , endocrinology
Lipid metabolism is spatiotemporally regulated within cells, yet intervention into lipid functions at subcellular resolution remains difficult. Here, we report a method that enables site-specific release of sphingolipids and cholesterol inside the vacuole in Saccharomyces cerevisiae . Using this approach, we monitored real-time sphingolipid metabolic flux out of the vacuole by mass spectrometry and found that the endoplasmic reticulum-vacuole-tethering protein Mdm1 facilitated the metabolism of sphingoid bases into ceramides. In addition, we showed that cholesterol, once delivered into yeast using our method, could restore cell proliferation induced by ergosterol deprivation, overcoming the previously described sterol-uptake barrier under aerobic conditions. Together, these data define a new way to study intracellular lipid metabolism and transport from the vacuole in yeast.