HIV-1 Rev mediates the nuclear export of intron-containing viral RNA transcripts and is essential for viral replication. Rev is imported into the nucleus by the host protein Importin beta (ImpB), but how Rev associates with ImpB is poorly understood. Here we report biochemical, biophysical and structural studies of the ImpB/Rev complex. Binding and mutagenesis data reveal that ImpB associates with two Rev monomers through independent binding sites and that the N-terminal half of Rev's Arginine-Rich Motif (ARM) is a primary ImpB binding epitope. Small-angle X-ray scattering (SAXS), crosslinking mass spectrometry and compensatory mutagenesis data suggest a structural model in which one Rev monomer binds to the C-terminal half of ImpB with Rev helix alpha-2 roughly parallel to the HEAT-repeat superhelical axis while the other monomer binds to the N-terminal half. These findings shed light on the molecular basis of Rev recognition by ImpB and highlight an atypical binding behaviour that distinguishes Rev from canonical cellular ImpB cargos.
Binding stoichiometry and structural model of the HIV-1 Rev/Importin beta complex
