PT  - JOURNAL ARTICLE
AU  - Takuma Kishimoto
AU  - Tetsuo Mioka
AU  - Eriko Itoh
AU  - David E. Williams
AU  - Raymond J. Andersen
AU  - Kazuma Tanaka
TI  - Phospholipid flippases and Sfk1 are essential for the retention of ergosterol in the plasma membrane
AID  - 10.1101/2020.10.19.346320
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.10.19.346320
4099  - http://biorxiv.org/content/early/2020/10/20/2020.10.19.346320.short
4100  - http://biorxiv.org/content/early/2020/10/20/2020.10.19.346320.full
AB  - Sterols are important lipid components of the plasma membrane (PM) in eukaryotic cells, but it is unknown how the PM retains sterols at a high concentration. Phospholipids are asymmetrically distributed in the PM, and phospholipid flippases play an important role in generating this phospholipid asymmetry. Here, we provide evidence that phospholipid flippases are essential for retaining ergosterol in the PM of yeast. A mutant in three flippases, Dnf1-Lem3, Dnf2-Lem3, and Dnf3-Crf1, and a membrane protein, Sfk1, showed a severe growth defect. We recently identified Sfk1 as a PM protein involved in phospholipid asymmetry. The PM of this mutant showed high permeability and low density, and many nutrient transporters failed to localize to the PM. Staining with the sterol probe filipin and the expression of a sterol biosensor revealed that ergosterol was not retained in the PM. Instead, ergosterol accumulated in an esterified form in lipid droplets. We propose that ergosterol is retained in the PM by the asymmetrical distribution of phospholipids and the action of Sfk1. Once phospholipid asymmetry is severely disrupted, sterols might be exposed on the cytoplasmic leaflet of the PM and actively transported to the endoplasmic reticulum by sterol transfer proteins.Competing Interest StatementThe authors have declared no competing interest.