title: Reconstitution of selective HIV-1 RNA packaging in vitro by membrane-bound Gag assemblies.
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.
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States.|California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, United States.
authors.forename: Lars-Anders
authors.initials: LA
authors.lastname: Carlson
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States.
authors.forename: Yun
authors.initials: Y
authors.lastname: Bai
authors.affiliation: Howard Hughes Medical Institute, Baltimore, United States.|Department of Chemistry and Biochemistry, University of Maryland Baltimore County, Baltimore, United States.
authors.forename: Sarah C
authors.initials: SC
authors.lastname: Keane
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States.|California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, United States.|Howard Hughes Medical Institute, University of California, Berkeley, Baltimore, United States.|Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, United States.
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Doudna
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States.|California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, United States.|Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, United States.
authors.forename: James H
authors.identifier: 0000-0001-5054-5445
authors.initials: JH
authors.lastname: Hurley
mesh_terms: D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D012367:RNA, Viral / Q000378:metabolism*; D053835:Unilamellar Liposomes / Q000378:metabolism*; D019065:Virus Assembly; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000378:metabolism*
chemical_list: D012367:RNA, Viral; D053835:Unilamellar Liposomes; D054301:gag Gene Products, Human Immunodeficiency Virus
keywords: HIV-1; SHAPE; biophysics; giant unilamellar vesicles; human; structural biology; viral genome packaging; virus assembly