An optogenetic system to control membrane phospholipid asymmetry through flippase activation in budding yeast

摘要: Lipid asymmetry in biological membranes is essential for various cell functions, such as polarity, cytokinesis, and apoptosis. P4-ATPases (flippases) are involved the generation of asymmetry. In Saccharomyces cerevisiae, protein kinases Fpk1p/Fpk2p activate Dnf1p/Dnf2p by phosphorylation. Previously, we have shown that a blue-light-dependent kinase, phototropin from Chlamydomonas reinhardtii (CrPHOT), complements defects an fpk1Δ fpk2Δ mutant. Herein, investigated whether CrPHOT optically regulates P4-ATPase activity. First, demonstrated translocation NBD-labelled phospholipids to cytoplasmic leaflet via was promoted blue-light irradiation cells with CrPHOT. addition, blue light completely suppressed membrane functions (such endocytic recycling, actin depolarization, apical-isotropic growth switching) caused mutations. All responses required kinase activity Hence, these results indicate utility powerful first tool optogenetic manipulation