Single-molecule imaging of HIV-1 envelope glycoprotein dynamics and Gag lattice association exposes determinants responsible for virus incorporation

Significance To create an infectious HIV-1 particle, the envelope glycoprotein (Env) must find a budding virus assembly site on the surface of an infected cell. Assembly sites are composed of growing 2-dimensional lattices of the structural Gag protein anchored to the plasma membrane by the matrix domain. The cytoplasmic tail of Env and the matrix domain are physically proximal on the inner leaflet of the membrane, placing these elements in direct contact upon encounter. We provide single-molecule evidence that the Env cytoplasmic tail and a single residue in matrix are necessary for lattice retention of Env, with membrane budding being dispensable. This supports a model for direct interaction or corralling of Env by the matrix domain and suggests that a druggable interface exists.
The HIV-1 envelope glycoprotein (Env) is sparsely incorporated onto assembling virus particles on the host cell plasma membrane in order for the virus to balance infectivity and evade the immune response. Env becomes trapped in a nascent particle on encounter with the polymeric viral protein Gag, which forms a dense protein lattice on the inner leaflet of the plasma membrane. While Env incorporation efficiency is readily measured biochemically from released particles, very little is known about the spatiotemporal dynamics of Env trapping events. Herein, we demonstrate, via high-resolution single-molecule tracking, that retention of Env trimers within single virus assembly sites requires the Env cytoplasmic tail (CT) and the L12 residue in the matrix (MA) domain of Gag but does not require curvature of the viral lattice. We further demonstrate that Env trimers are confined to subviral regions of a budding Gag lattice, supporting a model where direct interactions and/or steric corralling between the Env-CT and a lattice of MA trimers promote Env trapping and infectious HIV-1 assembly.