TY - JOUR
T1 - Vacuole-Specific Lipid Release for Tracking Intracellular Lipid Metabolism and Transport in Saccharomyces cerevisiae
AU - Girik, Vladimir
AU - Feng, Suihan
AU - Hariri, Hanaa
AU - Henne, W. Mike
AU - Riezman, Howard
N1 - Funding Information:
This work was supported by funds from the Welch Foundation (I-1873), the NIH NIGMS (GM119768), the NIDDK (R01DK126887), the Ara Parseghian Medical Research Fund, and the UT Southwestern Endowed Scholars Program (W.M.H.), and by the Leducq Foundation, the NCCR Chemical Biology and Swiss National Science Foundation (51NF40-185898 and 310030_184949) and the Canton of Geneva (H.R.).
Publisher Copyright:
© 2022 American Chemical Society. All rights reserved.
PY - 2022/6/17
Y1 - 2022/6/17
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85132455268&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85132455268&partnerID=8YFLogxK
U2 - 10.1021/acschembio.2c00120
DO - 10.1021/acschembio.2c00120
M3 - Article
C2 - 35667650
AN - SCOPUS:85132455268
SN - 1554-8929
VL - 17
SP - 1485
EP - 1494
JO - ACS chemical biology
JF - ACS chemical biology
IS - 6
ER -