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

Carlson, Lars-Anders; Bai, Yun; Keane, Sarah C.; Doudna, Jennifer A.; Hurley, James H.

doi:10.7554/elife.14663

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

Journal Article · Sat Jun 25 00:00:00 EDT 2016 · eLife

DOI:https://doi.org/10.7554/elife.14663· OSTI ID:1628851

Carlson, Lars-Anders ^[1]; Bai, Yun ^[2]; Keane, Sarah C. ^[3]; Doudna, Jennifer A. ^[4]; ^[5]

University of California, Berkeley, Berkeley (United States). Department of Molecular and Cell Biology; University of California, Berkeley, Berkeley (United States). California Institute for Quantitative Biosciences; DOE/OSTI
University of California, Berkeley, Berkeley (United States). Department of Molecular and Cell Biology
Howard Hughes Medical Institute, Baltimore (United States); University of Maryland Baltimore County, Baltimore (United States). Department of Chemistry and Biochemistry
University of California, Berkeley, Berkeley (United States). Department of Molecular and Cell Biology; University of California, Berkeley, Berkeley (United States). California Institute for Quantitative Bioscience; University of California, Berkeley, Baltimore (United States). Howard Hughes Medical Institute; Lawrence Berkeley National Laboratory, Berkeley (United States). Molecular Biophysics and Integrated Bioimaging Division
University of California, Berkeley, Berkeley (United States). Department of Molecular and Cell Biology; University of California, Berkeley, Berkeley (United States). California Institute for Quantitative Biosciences, Lawrence Berkeley National Laboratory, Berkeley (United States). Molecular Biophysics and Integrated Bioimaging Division

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.

View Accepted Manuscript (DOE)

Sponsoring Organization:: USDOE

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1628851

Journal Information:: eLife, Journal Name: eLife Vol. 5; ISSN 2050-084X

Publisher:: eLife Sciences Publications, Ltd.Copyright Statement

Country of Publication:: United States

Language:: English

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