Quantitative high-content imaging identifies novel regulators of Neo1 trafficking at endosomes.

摘要: P4-ATPases are a family of putative phospholipid flippases that regulate lipid membrane asymmetry, which is important for vesicle formation. Two yeast flippases, Drs2 and Neo1, have nonredundant functions in the recycling synaptobrevin-like v-SNARE Snc1 from early endosomes. activity needed to form vesicles its own trafficking, suggesting flippase localization linked. However, role Neo1 endosomal not well characterized. To identify novel regulators trafficking at endosomes, we first identified mutants with impaired Snc1-based reporter subsequently used high-content microscopy classify these based on or binding partners, Mon2 Dop1. This analysis Arl1 stabilizing Mon2/Dop1 complex uncovered new function Vps13 endosome localization. We further showed cargo-selective sorting nexin Snx3 required an motif N-terminus. Of importance, Snx3-dependent was correct another cargo protein, incorporation into tubules may influence their