Reconstitution of selective HIV-1 RNA packaging in vitro by membrane-bound Gag assemblies
Abstrak
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.
Topik & Kata Kunci
Penulis (5)
Lars-Anders Carlson
Yun Bai
Sarah C Keane
Jennifer A Doudna
James H Hurley
Akses Cepat
- Tahun Terbit
- 2016
- Sumber Database
- DOAJ
- DOI
- 10.7554/eLife.14663
- Akses
- Open Access ✓