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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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Main Authors: Müller, Thorsten G. (Author) , Zila, Vojtech (Author) , Peters, Kyra (Author) , Schifferdecker, Sandra (Author) , Stanić, Mia (Author) , Lucic, Bojana (Author) , Laketa, Vibor (Author) , Lusic, Marina (Author) , Müller, Barbara (Author) , Kräusslich, Hans-Georg (Author)
Format: Article (Journal)
Language:English
Published: Apr 27, 2021
In: eLife
Year: 2021, Volume: 10, Pages: 1-32
ISSN:2050-084X
DOI:10.7554/eLife.64776
Online Access:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.7554/eLife.64776
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Author Notes: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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Summary: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.
Item Description:Gesehen am 07.07.2021
Physical Description:Online Resource
ISSN:2050-084X
DOI:10.7554/eLife.64776

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