Imaging HIV-1 Nuclear Import, Uncoating, and Proviral Transcription.
Capsid
HIV-1
Live-cell imaging
Nuclear pore complex
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2024
2024
Historique:
medline:
15
5
2024
pubmed:
15
5
2024
entrez:
14
5
2024
Statut:
ppublish
Résumé
Live-cell imaging has become a powerful tool for dissecting the behavior of viral complexes during HIV-1 infection with high temporal and spatial resolution. Very few HIV-1 particles in a viral population are infectious and successfully complete replication (~1/50). Single-particle live-cell imaging enables the study of these rare infectious viral particles, which cannot be accomplished in biochemical assays that measure the average property of the entire viral population, most of which are not infectious. The timing and location of many events in the early stage of the HIV-1 life cycle, including nuclear import, uncoating, and integration, have only recently been elucidated. Live-cell imaging also provides a valuable approach to study interactions of viral and host factors in distinct cellular compartments and at specific stages of viral replication. Successful live-cell imaging experiments require careful consideration of the fluorescent labeling method used and avoid or minimize its potential impact on normal viral replication and produce misleading results. Ideally, it is beneficial to utilize multiple virus labeling strategies and compare the results to ensure that the virion labeling did not adversely influence the viral replication step that is under investigation. Another potential benefit of using different labeling strategies is that they can provide information about the state of the viral complexes. Here, we describe our methods that utilize multiple fluorescent protein labeling approaches to visualize and quantify important events in the HIV-1 life cycle, including docking HIV-1 particles with the nuclear envelope (NE) and their nuclear import, uncoating, and proviral transcription.
Identifiants
pubmed: 38743218
doi: 10.1007/978-1-0716-3862-0_2
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
15-30Informations de copyright
© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
Références
McDonald D, Vodicka MA, Lucero G, Svitkina TM, Borisy GG, Emerman M, Hope TJ (2002) Visualization of the intracellular behavior of HIV in living cells. J Cell Biol 159(3):441–452. https://doi.org/10.1083/jcb.200203150
doi: 10.1083/jcb.200203150
pubmed: 12417576
pmcid: 2173076
Chen J, Liu Y, Wu B, Nikolaitchik OA, Mohan PR, Chen J, Pathak VK, Hu WS (2020) Visualizing the translation and packaging of HIV-1 full-length RNA. Proc Natl Acad Sci USA 117(11):6145–6155. https://doi.org/10.1073/pnas.1917590117
doi: 10.1073/pnas.1917590117
pubmed: 32132202
pmcid: 7084099
Jouvenet N, Bieniasz PD, Simon SM (2008) Imaging the biogenesis of individual HIV-1 virions in live cells. Nature 454(7201):236–240. https://doi.org/10.1038/nature06998
doi: 10.1038/nature06998
pubmed: 18500329
pmcid: 2708942
Hubner W, Chen P, Del Portillo A, Liu Y, Gordon RE, Chen BK (2007) Sequence of human immunodeficiency virus type 1 (HIV-1) Gag localization and oligomerization monitored with live confocal imaging of a replication-competent, fluorescently tagged HIV-1. J Virol 81(22):12596–12607. https://doi.org/10.1128/JVI.01088-07
doi: 10.1128/JVI.01088-07
pubmed: 17728233
pmcid: 2168995
Chen J, Rahman SA, Nikolaitchik OA, Grunwald D, Sardo L, Burdick RC, Plisov S, Liang E, Tai S, Pathak VK, Hu WS (2016) HIV-1 RNA genome dimerizes on the plasma membrane in the presence of Gag protein. Proc Natl Acad Sci USA 113(2):E201–E208. https://doi.org/10.1073/pnas.1518572113
doi: 10.1073/pnas.1518572113
pubmed: 26712001
Hubner W, McNerney GP, Chen P, Dale BM, Gordon RE, Chuang FY, Li XD, Asmuth DM, Huser T, Chen BK (2009) Quantitative 3D video microscopy of HIV transfer across T cell virological synapses. Science 323(5922):1743–1747. https://doi.org/10.1126/science.1167525
doi: 10.1126/science.1167525
pubmed: 19325119
pmcid: 2756521
Albanese A, Arosio D, Terreni M, Cereseto A (2008) HIV-1 pre-integration complexes selectively target decondensed chromatin in the nuclear periphery. PLoS One 3(6):e2413. https://doi.org/10.1371/journal.pone.0002413
doi: 10.1371/journal.pone.0002413
pubmed: 18545681
pmcid: 2398779
Burdick RC, Li C, Munshi M, Rawson JMO, Nagashima K, Hu WS, Pathak VK (2020) HIV-1 uncoats in the nucleus near sites of integration. Proc Natl Acad Sci USA 117(10):5486–5493. https://doi.org/10.1073/pnas.1920631117
doi: 10.1073/pnas.1920631117
pubmed: 32094182
pmcid: 7071919
Francis AC, Melikyan GB (2018) Single HIV-1 imaging reveals progression of infection through CA-dependent steps of docking at the nuclear pore, uncoating, and nuclear transport. Cell Host Microbe 23(4):536–548.e536. https://doi.org/10.1016/j.chom.2018.03.009
doi: 10.1016/j.chom.2018.03.009
pubmed: 29649444
pmcid: 5901770
Schifferdecker S, Zila V, Muller TG, Sakin V, Anders-Osswein M, Laketa V, Krausslich HG, Muller B (2022) Direct capsid labeling of infectious HIV-1 by genetic code expansion allows detection of largely complete nuclear capsids and suggests nuclear entry of HIV-1 complexes via common routes. mBio 13(5):e0195922. https://doi.org/10.1128/mbio.01959-22
doi: 10.1128/mbio.01959-22
pubmed: 35972146
Arhel N, Genovesio A, Kim KA, Miko S, Perret E, Olivo-Marin JC, Shorte S, Charneau P (2006) Quantitative four-dimensional tracking of cytoplasmic and nuclear HIV-1 complexes. Nat Methods 3(10):817–824. https://doi.org/10.1038/nmeth928
doi: 10.1038/nmeth928
pubmed: 16990814
Muller TG, Zila V, Peters K, Schifferdecker S, Stanic M, Lucic B, Laketa V, Lusic M, Muller B, Krausslich HG (2021) HIV-1 uncoating by release of viral cDNA from capsid-like structures in the nucleus of infected cells. Elife 10:e64776. https://doi.org/10.7554/eLife.64776
doi: 10.7554/eLife.64776
pubmed: 33904396
pmcid: 8169111
Chen J, Nikolaitchik O, Singh J, Wright A, Bencsics CE, Coffin JM, Ni N, Lockett S, Pathak VK, Hu WS (2009) High efficiency of HIV-1 genomic RNA packaging and heterozygote formation revealed by single virion analysis. Proc Natl Acad Sci USA 106(32):13535–13540. https://doi.org/10.1073/pnas.0906822106
doi: 10.1073/pnas.0906822106
pubmed: 19628694
pmcid: 2714765
Francis AC, Marin M, Shi J, Aiken C, Melikyan GB (2016) Time-resolved imaging of single HIV-1 uncoating in vitro and in living cells. PLoS Pathog 12(6):e1005709. https://doi.org/10.1371/journal.ppat.1005709
doi: 10.1371/journal.ppat.1005709
pubmed: 27322072
pmcid: 4913920
Burdick RC, Hu WS, Pathak VK (2013) Nuclear import of APOBEC3F-labeled HIV-1 preintegration complexes. Proc Natl Acad Sci USA 110(49):E4780–E4789. https://doi.org/10.1073/pnas.1315996110
doi: 10.1073/pnas.1315996110
pubmed: 24248339
pmcid: 3856795
Li C, Burdick RC, Nagashima K, Hu WS, Pathak VK (2021) HIV-1 cores retain their integrity until minutes before uncoating in the nucleus. Proc Natl Acad Sci USA 118(10):e2019467118. https://doi.org/10.1073/pnas.2019467118
doi: 10.1073/pnas.2019467118
pubmed: 33649225
pmcid: 7958386
Burdick RC, Delviks-Frankenberry KA, Chen J, Janaka SK, Sastri J, Hu WS, Pathak VK (2017) Dynamics and regulation of nuclear import and nuclear movements of HIV-1 complexes. PLoS Pathog 13(8):e1006570. https://doi.org/10.1371/journal.ppat.1006570
doi: 10.1371/journal.ppat.1006570
pubmed: 28827840
pmcid: 5578721
Muller TG, Zila V, Muller B, Krausslich HG (2022) Nuclear capsid uncoating and reverse transcription of HIV-1. Annu Rev Virol 9(1):261–284. https://doi.org/10.1146/annurev-virology-020922-110929
doi: 10.1146/annurev-virology-020922-110929
pubmed: 35704745
De Clercq E, Yamamoto N, Pauwels R, Baba M, Schols D, Nakashima H, Balzarini J, Debyser Z, Murrer BA, Schwartz D et al (1992) Potent and selective inhibition of human immunodeficiency virus (HIV)-1 and HIV-2 replication by a class of bicyclams interacting with a viral uncoating event. Proc Natl Acad Sci USA 89(12):5286–5290. https://doi.org/10.1073/pnas.89.12.5286
doi: 10.1073/pnas.89.12.5286
pubmed: 1608936
pmcid: 49276
Unutmaz D, KewalRamani VN, Marmon S, Littman DR (1999) Cytokine signals are sufficient for HIV-1 infection of resting human T lymphocytes. J Exp Med 189(11):1735–1746. https://doi.org/10.1084/jem.189.11.1735
doi: 10.1084/jem.189.11.1735
pubmed: 10359577
pmcid: 2193071
Yee JK, Miyanohara A, LaPorte P, Bouic K, Burns JC, Friedmann T (1994) A general method for the generation of high-titer, pantropic retroviral vectors: highly efficient infection of primary hepatocytes. Proc Natl Acad Sci USA 91(20):9564–9568. https://doi.org/10.1073/pnas.91.20.9564
doi: 10.1073/pnas.91.20.9564
pubmed: 7937806
pmcid: 44853
Zenklusen D, Larson DR, Singer RH (2008) Single-RNA counting reveals alternative modes of gene expression in yeast. Nat Struct Mol Biol 15(12):1263–1271. https://doi.org/10.1038/nsmb.1514
doi: 10.1038/nsmb.1514
pubmed: 19011635
pmcid: 3154325
Smith JL, Pathak VK (2010) Identification of specific determinants of human APOBEC3F, APOBEC3C, and APOBEC3DE and African green monkey APOBEC3F that interact with HIV-1 Vif. J Virol 84(24):12599–12608. https://doi.org/10.1128/JVI.01437-10
doi: 10.1128/JVI.01437-10
pubmed: 20943965
pmcid: 3004357
Burdick R, Smith JL, Chaipan C, Friew Y, Chen J, Venkatachari NJ, Delviks-Frankenberry KA, Hu WS, Pathak VK (2010) P body-associated protein Mov10 inhibits HIV-1 replication at multiple stages. J Virol 84(19):10241–10253. https://doi.org/10.1128/JVI.00585-10
doi: 10.1128/JVI.00585-10
pubmed: 20668078
pmcid: 2937795
O’Doherty U, Swiggard WJ, Malim MH (2000) Human immunodeficiency virus type 1 spinoculation enhances infection through virus binding. J Virol 74(21):10074–10080. https://doi.org/10.1128/jvi.74.21.10074-10080.2000
doi: 10.1128/jvi.74.21.10074-10080.2000
pubmed: 11024136
pmcid: 102046
Holmes RK, Koning FA, Bishop KN, Malim MH (2007) APOBEC3F can inhibit the accumulation of HIV-1 reverse transcription products in the absence of hypermutation – comparisons with APOBEC3G. J Biol Chem 282(4):2587–2595. https://doi.org/10.1074/jbc.M607298200
doi: 10.1074/jbc.M607298200
pubmed: 17121840
Mbisa JL, Bu W, Pathak VK (2010) APOBEC3F and APOBEC3G inhibit HIV-1 DNA integration by different mechanisms. J Virol 84(10):5250–5259. https://doi.org/10.1128/Jvi.02358-09
doi: 10.1128/Jvi.02358-09
pubmed: 20219927
pmcid: 2863843