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Reconstitution of selective HIV-1 RNA packaging in vitro by membrane-bound Gag assemblies.

  • Author(s): Carlson, L-A
  • Bai, Y
  • Keane, SC
  • Doudna, JA
  • Hurley, JH
  • et al.

Published Web Location

https://elifesciences.org/content/5/e14663
No data is associated with this publication.
Abstract

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.

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