HIV-1 binds dynein directly to hijack microtubule transport machinery.

HIV-1 uses the microtubule cytoskeleton to reach the host cell nucleus during replication, yet the molecular basis for microtubule-dependent HIV-1 motility is poorly understood. Using reconstitution biochemistry and single-molecule imaging, we found that HIV-1 binds to the retrograde microtubule-associated motor, dynein, directly and not via a cargo adaptor, as has been previously suggested. We found dynein's tail domain binds to a novel interface on the HIV-1 capsid lattice. Further, we use an engineered encapsulin system to demonstrate that multiple dynein motors tethered to rigid cargoes, like HIV-1 capsids, display reduced motility, distinct from the behavior of multiple motors on membraneous cargoes. Our results provide a new model of HIV-1 trafficking wherein HIV-1 'hijacks' the dynein transport machinery for microtubule motility, establishing a new model of viral trafficking by directly co-opting host dynein motors.