HIV-1 uncoating by release of viral cDNA from capsid-like structures in the nucleus of infected cells
HIV-1 replication commences inside the cone-shaped viral capsid, but timing, localization, and mechanism of uncoating are under debate. We adapted a strategy to visualize individual reverse-transcribed HIV-1 cDNA molecules and their association with viral and cellular proteins using fluorescence and...
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| Hauptverfasser: | , , , , , , , , , |
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| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
Apr 27, 2021
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| In: |
eLife
Year: 2021, Jahrgang: 10, Pages: 1-32 |
| ISSN: | 2050-084X |
| DOI: | 10.7554/eLife.64776 |
| Online-Zugang: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.7554/eLife.64776
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| Verfasserangaben: | Thorsten G Müller, Vojtech Zila, Kyra Peters, Sandra Schifferdecker, Mia Stanic, Bojana Lucic, Vibor Laketa, Marina Lusic, Barbara Müller, Hans-Georg Kräusslich |
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| 520 | |a HIV-1 replication commences inside the cone-shaped viral capsid, but timing, localization, and mechanism of uncoating are under debate. We adapted a strategy to visualize individual reverse-transcribed HIV-1 cDNA molecules and their association with viral and cellular proteins using fluorescence and correlative-light-and-electron-microscopy (CLEM). We specifically detected HIV-1 cDNA inside nuclei, but not in the cytoplasm. Nuclear cDNA initially co-localized with a fluorescent integrase fusion (IN-FP) and the viral CA (capsid) protein, but cDNA-punctae separated from IN-FP/CA over time. This phenotype was conserved in primary HIV-1 target cells, with nuclear HIV-1 complexes exhibiting strong CA-signals in all cell types. CLEM revealed cone-shaped HIV-1 capsid-like structures and apparently broken capsid-remnants at the position of IN-FP signals and elongated chromatin-like structures in the position of viral cDNA punctae lacking IN-FP. Our data argue for nuclear uncoating by physical disruption rather than cooperative disassembly of the CA-lattice, followed by physical separation from the pre-integration complex. | ||
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