Reconstitution of selective HIV-1 RNA packaging in vitro by membrane-bound Gag assemblies.
- Author(s): Carlson, Lars-Anders
- Bai, Yun
- Keane, Sarah C
- Doudna, Jennifer A
- Hurley, James H
- et al.
Published Web Locationhttps://elifesciences.org/content/5/e14663
HIV-1 Gag selects and packages a dimeric, unspliced viral RNA in the context of a large excess of cytosolic human RNAs. As Gag assembles on the plasma membrane, the HIV-1 genome is enriched relative to cellular RNAs by an unknown mechanism. We used a minimal system consisting of purified RNAs, recombinant HIV-1 Gag and giant unilamellar vesicles to recapitulate the selective packaging of the 5' untranslated region of the HIV-1 genome in the presence of excess competitor RNA. Mutations in the CA-CTD domain of Gag which subtly affect the self-assembly of Gag abrogated RNA selectivity. We further found that tRNA suppresses Gag membrane binding less when Gag has bound viral RNA. The ability of HIV-1 Gag to selectively package its RNA genome and its self-assembly on membranes are thus interdependent on one another.