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Complementary assays reveal a relationship between HIV-1 uncoating and reverse transcription
Open accessAbstract
During the early stages of HIV-1 replication the conical capsid composed of p24 CA protein dissociates from the rest of the cytoplasmic viral complex by a process called uncoating. Although proper uncoating is known to be required for HIV-1 infection, many questions remain about the timing and factors involved in the process. Here we have used two complementary assays to study the process of uncoating in HIV-1–infected cells, specifically looking at the timing of uncoating and its relationship to reverse transcription. We developed a fluorescent microscopy-based uncoating assay that detects the association of p24 CA with HIV-1 viral complexes in cells. We also used an owl monkey kidney (OMK) cell assay that is based on timed TRIM-CypA–mediated restriction of HIV-1 replication. Results from both assays indicate that uncoating is initiated within 1 h of viral fusion. In addition, treatment with the reverse transcriptase inhibitor nevirapine delayed uncoating in both assays. Analysis of reverse transcription products in OMK cells revealed that the generation of early reverse transcription products coincides with the timing of uncoating in these assays. Collectively, these results suggest that some aspect of reverse transcription has the ability to influence the kinetics of uncoating.
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“This RTC is actively transported to the chromosomal DNA, where integration may occur [6–9].”
From Clinical use of lentiviral vectors (2018) · cites this paper as [9]
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Pak, A. J., Grime, J. M. A., Yu, A., & Voth, G. A. (2019) · Journal of the American Chemical Society · doi:10.1021/jacs.9b01413 · Cited by 50
HIV-1 requires capsid remodelling at the nuclear pore for nuclear entry and integration
Guedán, A., Donaldson, C. D., Caroe, E. R., Cosnefroy, O., Taylor, I. A., & Bishop, K. N. (2021) · PLoS Pathogens · doi:10.1371/journal.ppat.1009484 · Cited by 47
Bulli, L., Apolonia, L., Kutzner, J., Pollpeter, D., Goujon, C., Herold, N., Schwarz, S.-M., Giernat, Y., Keppler, O. T., Malim, M. H., et al. (2016) · Journal of Virology · doi:10.1128/jvi.00458-16 · Cited by 46
Recognition of HIV-1 capsid by PQBP1 licenses an innate immune sensing of nascent HIV-1 DNA
Yoh, S. M., Mamede, J. I., Lau, D., Ahn, N., Sánchez-Aparicio, M. T., Temple, J. E., Tuckwell, A., Fuchs, N. V., Cianci, G. C., Riva, L., et al. (2022) · Molecular Cell · doi:10.1016/j.molcel.2022.06.010 · Cited by 47
Self-citationImaging HIV-1 Genomic DNA from Entry through Productive Infection
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Siddiqui, M. A., Saito, A., Halambage, U. D., Ferhadian, D., Fischer, D. K., Francis, A. C., Melikyan, G. B., Ambrose, Z., Aiken, C., & Yamashita, M. (2019) · Journal of Virology · doi:10.1128/jvi.00706-19 · Cited by 42
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Capsid-Labelled HIV To Investigate the Role of Capsid during Nuclear Import and Integration
Zurnic, I., Dirix, L., Lemmens, V., Borrenberghs, D., Wit, F. D., Vernaillen, F., Rocha, S., Christ, F., Hendrix, J., Hofkens, J., et al. (2020) · Journal of Virology · doi:10.1128/jvi.01024-19 · Cited by 44
The capsid lattice engages a bipartite NUP153 motif to mediate nuclear entry of HIV-1 cores
Shen, Q., Kumari, S., Xu, C., Jang, S., Shi, J., Burdick, R. C., Levintov, L., Xiong, Q., Wu, C., Devarkar, S. C., et al. (2023) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.2202815120 · Cited by 41
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HIV-1 uncoating requires long double-stranded reverse transcription products
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Metadata
- Authors
- Hulme, A. E., Perez, O., & Hope, T. J.
- Affiliations
- Northwestern University (US)
- University of Illinois Chicago (US)
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2011-05-31
- Volume
- 108
- Issue
- 24
- Pages
- 9975-9980
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC3116424
- OpenAlex
- W2120792521
- Cited by
- 276
- References
- 29
- Citations by year
- 2026: 6 · 2025: 8 · 2024: 14 · 2023: 9 · 2022: 18 · 2021: 24 · 2020: 24 · 2019: 20 · 2018: 23 · 2017: 15 · 2016: 19 · 2015: 15 · 2014: 24 · 2013: 27 · 2012: 22
- FWCI
- 10.25
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- BearWorks (Missouri State University) (submittedVersion)
- Funders
- National Institute of General Medical Sciences (P50GM082545)
- National Institute of Allergy and Infectious Diseases (R01-AI47770, R01AI047770)
- Topics
- Life Sciences › Immunology and Microbiology › Virology › HIV Research and Treatment
- Health Sciences › Medicine › Infectious Diseases › HIV/AIDS drug development and treatment
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and Retrovirus Studies
- Keywords
- reverse transcription, viral capsid, viral fusion, nevirapine
- MeSH terms
- Animals, Cell Line, HeLa Cells, Humans, Luminescent Proteins, Microscopy, Fluorescence, Recombinant Fusion Proteins, Time Factors, Virion, Virus Replication, HIV-1, Cyclosporine, HIV Core Protein p24, Macrolides, Anti-HIV Agents, Nevirapine, Reverse Transcription, Green Fluorescent Proteins, Human Immunodeficiency Virus Proteins, vpr Gene Products, Human Immunodeficiency Virus, HEK293 Cells
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