Specific Interactions between HIV-1 Env Cytoplasmic Tail and Gag Matrix Domain Probed by NMR

Abstract

HIV-1 envelope glycoprotein (Env) is a transmembrane protein that mediates membrane fusion during viral entry. Incorporation of a sufficient number of Envs during viral assembly is critical for viral infectivity. It has long been suggested that the interaction between Env and the matrix domain (MA) of the Gag polyprotein plays an important role in recruiting Envs to the site of viral assembly on the plasma membrane, but direct biochemical and structural evidence is lacking for such an interaction in the context of a membrane-like environment. Here, we report specific structural contacts between the cytoplasmic tail (CT) of the trimeric HIV-1 Env in bicelles and the trimeric MA. Using a combination of measurements of NMR chemical shift perturbation, intermolecular paramagnetic relaxation enhancements, and microscale thermophoresis, we found that, in DMPC-DHPC bicelles that mimic a lipid bilayer, the trimeric baseplate formed by the CT specifically interacted with the trimeric MA via mostly electrostatic interactions involving acidic residues of the CT and positively charged patches of the MA. Nonconservative substitution of these previously unrecognized acidic residues in Env resulted in drastically reduced viral infectivity. Our findings, together with early genetic and biochemical studies, indicate that specific interactions between the CT of Env and MA play a structural role during HIV-1 assembly.