期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2022
卷号：119
期号：2
DOI：10.1073/pnas.2120411119
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：Significance
Cholesterol constitutes 50% of lipids in the plasma membrane (PM) of animal cells. Sensors in the endoplasmic reticulum (ER) maintain this level by adjusting cholesterol uptake, synthesis, and storage. Uptake is mediated by LDL receptors, which deliver cholesterol-carrying LDL to lysosomes from which cholesterol moves to the PM and then to the ER. We report PM-to-ER transport of LDL cholesterol requires cholesterol-binding Aster proteins anchored to the ER and phosphatidylserine embedded in the PM. Asters are known to bind phosphatidylserine, and this accounts for part of the phosphatidylserine requirement. However, the current data suggest an additional requirement for phosphatidylserine independent of Asters. These data advance our knowledge of PM cholesterol homeostasis, a control mechanism essential for cell growth and survival.
Low-density lipoprotein (LDL) delivers cholesterol to mammalian cells through receptor-mediated endocytosis. The LDL cholesterol is liberated in lysosomes and transported to the plasma membrane (PM) and from there to the endoplasmic reticulum (ER). Excess ER cholesterol is esterified with a fatty acid for storage as cholesteryl esters. Recently, we showed that PM-to-ER transport of LDL cholesterol requires phosphatidylserine (PS). Others showed that PM-to-ER transport of cholesterol derived from other sources requires Asters (also called GRAMD1s), a family of three ER proteins that bridge between the ER and PM by binding to PS. Here, we use a cholesterol esterification assay and other measures of ER cholesterol delivery to demonstrate that Asters participate in PM-to-ER transport of LDL cholesterol in Chinese hamster ovary cells. Knockout of the gene encoding PTDSS1, the major PS-synthesizing enzyme, lowered LDL-stimulated cholesterol esterification by 85%, whereas knockout of all three Aster genes lowered esterification by 65%. The reduction was even greater (94%) when the genes encoding PTDSS1 and the three Asters were knocked out simultaneously. We conclude that Asters participate in LDL cholesterol delivery from PM to ER, and their action depends in large part, but not exclusively, on PS. The data also indicate that PS participates in another delivery pathway, so far undefined, that is independent of Asters.
