20 p.-8 fig.
Low-density lipoprotein (LDL)-cholesterol delivery from late endosomes to the plasma membrane regulates focal adhesion dynamics and cell migration, but the mechanisms controlling it are poorly characterized. Here, we employed auxin-inducible rapid degradation of oxysterol-binding protein-related protein 2 (ORP2/OSBPL2) to show that endogenous ORP2 mediates the transfer of LDL-derived cholesterol from late endosomes to focal adhesion kinase (FAK)-/integrin-positive recycling endosomes in human cells. In vitro, cholesterol enhances membrane association of FAK to PI(4,5)P2-containing lipid bilayers. In cells, ORP2 stimulates FAK activation and PI(4,5)P2 generation in endomembranes, enhancing cell adhesion. Moreover, ORP2 increases PI(4,5)P2 in NPC1-containing late endosomes in a FAK-dependent manner, controlling their tubulovesicular trafficking. Together, these results provide evidence that ORP2 controls FAK activation and LDL-cholesterol plasma membrane delivery by promoting bidirectional cholesterol/PI(4,5)P2 exchange between late and recycling endosomes.
This study was supported by the Academy of Finland (grants 307415 and 324929 to E.I., 322647 to V.M.O.), Sigrid Juselius Foundation (E.I., V.M.O., L. A.-S.), Fondation Leducq (grant 19CVD04) (E.I.), Jane and Aatos Erkko Foundation (E.I.), Magnus Ehrnrooth Foundation (K.T., V.M.O.), and the Finnish Foundation for Cardiovascular Research (V.M.O.). L.-A.S. acknowledges support from Helsinki Institute of Life Science (HiLIFE, start-up grant) and D.L. from the Ministry of Science, Innovation and Universities for the Spanish State Research Agency Retos Grant RTI2018-099318-B-I00, cofounded by the European Regional Development Fund (FEDER).