Goodale, A.; Huang, S.-W.; Almeida, N.; Williamson, D.; Shkriabai, N.; Betancor, G.; Apolonia, L.; DiMaio, F.; Padilla-Parra, S.; Kvaratskhelia, M.; Bergeron, J.; Malim, M. M.

The HIV-1 capsid core encapsulates the viral genome and mediates its delivery into the nucleus of the host cell. It is composed of multiple copies of the Capsid (CA, p24Gag) protein, assembled into hexamers and pentamers to create a lattice that forms a fullerene-like cone. Myxovirus resistance 2 (MX2) is an HIV-1 restriction factor that binds to the capsid core and blocks nuclear import of the viral genome. Here, we define a minimal region of MX2 required for HIV-1 restriction and produce a corresponding functional recombinant protein. We have used cryo-electron microscopy to determine the structure of this MX2 fragment bound to the tri-hexamer interface of the capsid lattice, revealing a large, buried interface combining electrostatic and hydrophobic interactions. This structure, together with assays that measure capsid core destabilisation, shows that MX2 binding induces conformational rearrangements in the capsid lattice that culminate in a loss of integrity. These results support a model whereby MX2 exerts its antiviral activity by disrupting the capsid lattice, inducing premature fragmentation and preventing HIV-1 nuclear import. By revealing the structural basis for MX2-mediated restriction, this work also provides the framework for the development of anti-HIV molecules that mimic MX2 restriction.