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

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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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Lars-Anders Carlson

Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States

Competing interests
The authors declare that no competing interests exist.
Yun Bai

School of Life Science and Technology, ShanghaiTech University, Shanghai, China

Competing interests
The authors declare that no competing interests exist.
Sarah C Keane

Howard Hughes Medical Institute, University of California, Berkeley, Baltimore, United States

Competing interests
The authors declare that no competing interests exist.
Jennifer A Doudna

Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States

Competing interests
The authors declare that no competing interests exist.
James H Hurley

Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States

For correspondence
jimhurley@berkeley.edu

Competing interests
The authors declare that no competing interests exist.

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