Model showing control of Rab7–RILP–p150^Glued motor protein complexes by the ER protein VAP, cholesterol, and the sensor ORP1L. Rab7 recruits the homodimeric effector protein RILP to LEs. RILP binds the p150^Glued subunit of the dynein–dynactin motor. ORP1L also binds to the Rab7–RILP complex. ORP1L has a C-terminal cholesterol-sensing ORD that exists in different conformational states determined by cholesterol in LEs. At low cholesterol levels, ORD adopts a conformation in which the adjacent FFAT motif is exposed and which can be detected by the ER protein VAP in ER–LE MCSs. VAP binds in trans to the Rab7–RILP–p150^Glued complex and removes p150^Glued, thus preventing minus end–directed transport. High cholesterol conditions initiate a different ORP1L conformation, preventing formation of ER–LE MCSs, and VAP fails to interact with the Rab7–RILP–p150^Glued complex. Thus, cholesterol levels in LEs determine the conformation of ORP1L and thereby VAP recruitment in ER–LE contact sites. The ER protein VAP then controls p150^Glued binding to Rab7–RILP, resulting in the scattering of cholesterol-poor LEs and clustering of cholesterol-laden LEs, as in Niemann-Pick type C disease.