+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-11967 | |||||||||
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Title | Human nuclear pore complex in HIV-1 infected T cellsNuclear pore | |||||||||
Map data | Human Nuclear Pore Complex from HIV-1 infected T cellsNuclear pore | |||||||||
Sample |
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Biological species | Homo sapiens (human) | |||||||||
Method | subtomogram averaging / cryo EM / Resolution: 37.0 Å | |||||||||
Authors | Margiotta E / Beck M | |||||||||
Funding support | Germany, 1 items
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Citation | Journal: Cell / Year: 2021 Title: Cone-shaped HIV-1 capsids are transported through intact nuclear pores. Authors: Vojtech Zila / Erica Margiotta / Beata Turoňová / Thorsten G Müller / Christian E Zimmerli / Simone Mattei / Matteo Allegretti / Kathleen Börner / Jona Rada / Barbara Müller / Marina ...Authors: Vojtech Zila / Erica Margiotta / Beata Turoňová / Thorsten G Müller / Christian E Zimmerli / Simone Mattei / Matteo Allegretti / Kathleen Börner / Jona Rada / Barbara Müller / Marina Lusic / Hans-Georg Kräusslich / Martin Beck / Abstract: Human immunodeficiency virus (HIV-1) remains a major health threat. Viral capsid uncoating and nuclear import of the viral genome are critical for productive infection. The size of the HIV-1 capsid ...Human immunodeficiency virus (HIV-1) remains a major health threat. Viral capsid uncoating and nuclear import of the viral genome are critical for productive infection. The size of the HIV-1 capsid is generally believed to exceed the diameter of the nuclear pore complex (NPC), indicating that capsid uncoating has to occur prior to nuclear import. Here, we combined correlative light and electron microscopy with subtomogram averaging to capture the structural status of reverse transcription-competent HIV-1 complexes in infected T cells. We demonstrated that the diameter of the NPC in cellulo is sufficient for the import of apparently intact, cone-shaped capsids. Subsequent to nuclear import, we detected disrupted and empty capsid fragments, indicating that uncoating of the replication complex occurs by breaking the capsid open, and not by disassembly into individual subunits. Our data directly visualize a key step in HIV-1 replication and enhance our mechanistic understanding of the viral life cycle. #1: Journal: To Be Published Title: Cone-shaped HIV-1 capsids are transported through intact nuclear pores Authors: Zila V / Margiotta E | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_11967.map.gz | 2.5 MB | EMDB map data format | |
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Header (meta data) | emd-11967-v30.xml emd-11967.xml | 9.4 KB 9.4 KB | Display Display | EMDB header |
Images | emd_11967.png | 40.5 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-11967 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-11967 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_11967.map.gz / Format: CCP4 / Size: 11.4 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Human Nuclear Pore Complex from HIV-1 infected T cells | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 13.8 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : Nuclear pore complex from HIV-1 infected T cells
Entire | Name: Nuclear pore complex from HIV-1 infected T cellsNuclear pore |
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Components |
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-Supramolecule #1: Nuclear pore complex from HIV-1 infected T cells
Supramolecule | Name: Nuclear pore complex from HIV-1 infected T cells / type: complex / ID: 1 / Parent: 0 |
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Source (natural) | Organism: Homo sapiens (human) |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | subtomogram averaging |
Aggregation state | cell |
-Sample preparation
Buffer | pH: 7.4 / Details: RPMI 1640 medium with GlutaMAX |
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Grid | Model: Quantifoil / Material: GOLD / Mesh: 200 / Pretreatment - Type: GLOW DISCHARGE |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 310.15 K / Instrument: LEICA EM GP |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: OTHER / Imaging mode: OTHER / Nominal defocus max: 5.0 µm / Nominal defocus min: 2.0 µm / Nominal magnification: 42000 |
Image recording | Film or detector model: GATAN K2 QUANTUM (4k x 4k) / Average electron dose: 4.0 e/Å2 |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Extraction | Number tomograms: 250 / Number images used: 99 |
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CTF correction | Details: novaCTF (Turonova et al., 2017) |
Final angle assignment | Type: PROJECTION MATCHING |
Final reconstruction | Applied symmetry - Point group: C8 (8 fold cyclic) / Resolution.type: BY AUTHOR / Resolution: 37.0 Å / Resolution method: FSC 0.143 CUT-OFF / Number subtomograms used: 90 |