Reconstitution of selective HIV-1 RNA packaging in vitro by membrane-bound Gag assemblies
Open accessAbstract
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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The inhibitory effect of tRNA on Gag-membrane binding was validated in a minimal system including giant unilamellar vesicles, recombinant HIV-1 Gag protein, and purified RNAs.
“The inhibitory effect of tRNA on Gag-membrane binding was further validated in a minimal system that includes giant unilamellar vesicles, recombinant HIV-1 Gag protein, and purified RNAs (82).”
From Role of host tRNAs and aminoacyl-tRNA synthetases in retroviral replication (2019) · cites this paper as [82]
A model holds that viral gRNA-bound Gag overcomes the inhibitory effect of tRNA and thereby multimerizes and clusters at the plasma membrane.
“These observations led to another model wherein viral gRNA-bound Gag overcomes the inhibitory effect of tRNA and thereby multimerizes and clusters at the plasma membrane (Fig. 3B) (82).”
From Role of host tRNAs and aminoacyl-tRNA synthetases in retroviral replication (2019) · cites this paper as [82]
In the presence of tRNA, gRNA-bound Gag exhibits a higher likelihood of membrane binding than nonspecific RNA-bound Gag.
“In the presence of tRNA, gRNA-bound Gag exhibits a higher likelihood of membrane binding than nonspecific RNA-bound Gag (82).”
From Role of host tRNAs and aminoacyl-tRNA synthetases in retroviral replication (2019) · cites this paper as [82]
tRNA plays a regulatory role in Gag membrane binding.
“tRNA plays a regulatory role in Gag membrane binding.”
From Role of host tRNAs and aminoacyl-tRNA synthetases in retroviral replication (2019) · cites this paper as [82]
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Role of host tRNAs and aminoacyl-tRNA synthetases in retroviral replication
Jin, D., & Musier-Forsyth, K. (2019) · Journal of Biological Chemistry · doi:10.1074/jbc.rev118.002957 · Cited by 67 · see citing works
- Authors
- Carlson, L.-A., Bai, Y., Keane, S. C., Doudna, J. A., & Hurley, J. H.
- Affiliations
- QB3 (US)
- University of California, Berkeley (US)
- Howard Hughes Medical Institute (US)
- Corresponding author
- Carlson, Lars-Anders
- Hurley, James H
- Venue
- eLife
- Publisher
- eLife Sciences Publications, Ltd
- Published
- 2016-06-25
- Volume
- 5
- ISSN
- 2050-084X (ISSN-L)
- PMC
- PMC4946900
- OpenAlex
- W2461247482
- Cited by
- 48
- References
- 6
- Citations by year
- 2026: 6 · 2025: 1 · 2024: 5 · 2023: 4 · 2022: 6 · 2021: 3 · 2020: 3 · 2019: 5 · 2018: 7 · 2017: 6 · 2016: 2
- FWCI
- 4.62
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- eLife (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- OSTI OAI (U.S. Department of Energy Office of Scientific and Technical Information) (submittedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- eScholarship (California Digital Library) (submittedVersion)
- Funders
- National Institute of Allergy and Infectious Diseases (R01AI112442)
- National Institute of General Medical Sciences (P50GM082250, P50GM103297)
- Topics
- Life Sciences › Immunology and Microbiology › Virology › HIV Research and Treatment
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and Retrovirus Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Keywords
- HIV-1, RNA packaging, tRNA, 5' untranslated region, unspliced viral RNA, in vitro reconstitution, giant unilamellar vesicles
- MeSH terms
- Humans, RNA, Viral, HIV-1, Virus Assembly, Unilamellar Liposomes, gag Gene Products, Human Immunodeficiency Virus
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