Updated project metadata. Timely uncoating and nuclear import are key to efficient HIV infection, however the triggers and mechanisms that orchestrate these steps are unknown. Here we show that HIV-1 exploits Transportin-1/TRN-1 for both uncoating and nuclear import. Depletion of TRN-1, which we characterised by mass spectrometry, significantly reduced the early steps of HIV-1 infection in cell lines and primary CD4+ T cells. We showed that TRN-1 binds to incoming HIV-1 capsid (CA) cores, but not monomeric CA or integrase, via a NLS present on the cyclophilin A (CypA) binding loop. The G89V mutant, but not P90A, relieved dependency on TRN-1, and TRN-1 binding to CA at position G89 displaced CypA, which likely contributes to weakening the CA assembly. Recombinant TRN-1 could induce CA uncoating in vitro indicating that disruption is entirely mechanical, and activity mapped to residue W730 that plays a pivotal role in binding to substrate NLS. HIV-1 intercepts TRN-1 near the nuclear envelope and complexes that fail to do so scatter back into the cytoplasm without uncoating. For complexes that uncoat, TRN-1 mediates the concomitant nuclear import of CA and viral DNA. Our study reveals how HIV-1 uses a single cellular protein to orchestrate the key steps that allow it to reach its replication compartment efficiently.