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- PDB-5md3: The structure of the mature HIV-1 CA hexameric lattice with curva... -

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Basic information

Entry
Database: PDB / ID: 5md3
TitleThe structure of the mature HIV-1 CA hexameric lattice with curvature parameters: tilt=11, twist=12
Components(Capsid protein p24) x 2
KeywordsVIRAL PROTEIN / retrovirus / HIV-1 / capsid / lattice curvature
Function / homology
Function and homology information


viral budding via host ESCRT complex / viral process / viral capsid / viral nucleocapsid / host cell cytoplasm / viral translational frameshifting / host cell nucleus / structural molecule activity / virion membrane / RNA binding ...viral budding via host ESCRT complex / viral process / viral capsid / viral nucleocapsid / host cell cytoplasm / viral translational frameshifting / host cell nucleus / structural molecule activity / virion membrane / RNA binding / zinc ion binding / cytoplasm
Similarity search - Function
Gag protein p6 / Gag protein p6 / : / gag protein p24 N-terminal domain / Immunodeficiency lentiviral matrix, N-terminal / gag gene protein p17 (matrix protein) / Matrix protein, lentiviral and alpha-retroviral, N-terminal / Retroviral nucleocapsid Gag protein p24, C-terminal domain / Gag protein p24 C-terminal domain / Retrovirus capsid, C-terminal ...Gag protein p6 / Gag protein p6 / : / gag protein p24 N-terminal domain / Immunodeficiency lentiviral matrix, N-terminal / gag gene protein p17 (matrix protein) / Matrix protein, lentiviral and alpha-retroviral, N-terminal / Retroviral nucleocapsid Gag protein p24, C-terminal domain / Gag protein p24 C-terminal domain / Retrovirus capsid, C-terminal / Retroviral matrix protein / Retrovirus capsid, N-terminal / zinc finger / Zinc knuckle / Zinc finger, CCHC-type superfamily / Zinc finger, CCHC-type / Zinc finger CCHC-type profile.
Similarity search - Domain/homology
Gag protein / Gag protein / Gag polyprotein
Similarity search - Component
Biological speciesHuman immunodeficiency virus 1
MethodELECTRON MICROSCOPY / subtomogram averaging / cryo EM / Resolution: 8.5 Å
AuthorsMattei, S. / Glass, B. / Hagen, W.J.H. / Kraeusslich, H.-G. / Briggs, J.A.G.
Funding support Germany, 2items
OrganizationGrant numberCountry
German Research FoundationBR 3635/2-1 Germany
German Research FoundationKR 906/7-1 Germany
CitationJournal: Science / Year: 2016
Title: The structure and flexibility of conical HIV-1 capsids determined within intact virions.
Authors: Simone Mattei / Bärbel Glass / Wim J H Hagen / Hans-Georg Kräusslich / John A G Briggs /
Abstract: HIV-1 contains a cone-shaped capsid encasing the viral genome. This capsid is thought to follow fullerene geometry-a curved hexameric lattice of the capsid protein, CA, closed by incorporating 12 CA ...HIV-1 contains a cone-shaped capsid encasing the viral genome. This capsid is thought to follow fullerene geometry-a curved hexameric lattice of the capsid protein, CA, closed by incorporating 12 CA pentamers. Current models for core structure are based on crystallography of hexameric and cross-linked pentameric CA, electron microscopy of tubular CA arrays, and simulations. Here, we report subnanometer-resolution cryo-electron tomography structures of hexameric and pentameric CA within intact HIV-1 particles. Whereas the hexamer structure is compatible with crystallography studies, the pentamer forms using different interfaces. Determining multiple structures revealed how CA flexes to form the variably curved core shell. We show that HIV-1 CA assembles both aberrant and perfect fullerene cones, supporting models in which conical cores assemble de novo after maturation.
History
DepositionNov 10, 2016Deposition site: PDBE / Processing site: PDBE
Revision 1.0Dec 28, 2016Provider: repository / Type: Initial release
Revision 1.1Aug 30, 2017Group: Author supporting evidence / Data collection ...Author supporting evidence / Data collection / Experimental preparation / Refinement description
Category: em_3d_fitting / em_imaging_optics ...em_3d_fitting / em_imaging_optics / em_sample_support / em_software / pdbx_audit_support
Item: _em_3d_fitting.target_criteria / _em_imaging_optics.energyfilter_name ..._em_3d_fitting.target_criteria / _em_imaging_optics.energyfilter_name / _em_sample_support.grid_type / _em_software.name / _pdbx_audit_support.funding_organization
Revision 1.2May 15, 2024Group: Data collection / Database references / Category: chem_comp_atom / chem_comp_bond / database_2
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession

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Assembly

Deposited unit
A: Capsid protein p24
B: Capsid protein p24
F: Capsid protein p24
W: Capsid protein p24
a: Capsid protein p24
b: Capsid protein p24
f: Capsid protein p24
C: Capsid protein p24
D: Capsid protein p24
E: Capsid protein p24
G: Capsid protein p24
H: Capsid protein p24
I: Capsid protein p24
J: Capsid protein p24


Theoretical massNumber of molelcules
Total (without water)164,77514
Polymers164,77514
Non-polymers00
Water00
1


  • Idetical with deposited unit
  • defined by author
TypeNameSymmetry operationNumber
identity operation1_5551

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Components

#1: Protein
Capsid protein p24


Mass: 8370.568 Da / Num. of mol.: 8 / Fragment: C-terminal domain, UNP residues 141-214 / Source method: isolated from a natural source / Source: (natural) Human immunodeficiency virus 1 / Cell line: MT-4 / Strain: NL43 / References: UniProt: A0A0K0V8J8, UniProt: Q72497*PLUS
#2: Protein
Capsid protein p24


Mass: 16301.689 Da / Num. of mol.: 6 / Fragment: N-terminal domain, UNP residues 123-269 / Source method: isolated from a natural source / Source: (natural) Human immunodeficiency virus 1 / Cell line: MT-4 / Strain: NL43 / References: UniProt: E9MJT0, UniProt: Q72497*PLUS

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Experimental details

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Experiment

ExperimentMethod: ELECTRON MICROSCOPY
EM experimentAggregation state: PARTICLE / 3D reconstruction method: subtomogram averaging

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Sample preparation

ComponentName: Human immunodeficiency virus 1 / Type: VIRUS
Details: HIV-1 particles were produced by infection of MT-4 cells with HIV-1 strain NL43 by coculture.
Entity ID: all / Source: NATURAL
Source (natural)Organism: Human immunodeficiency virus 1
Details of virusEmpty: NO / Enveloped: YES / Isolate: STRAIN / Type: VIRION
Natural hostOrganism: Homo sapiens
Buffer solutionpH: 7.4
Buffer componentName: PBS
SpecimenEmbedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
Details: Viral particles were purified from cell culture supernatant by iodixanol gradient centrifugation.
Specimen supportGrid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: C-flat
VitrificationInstrument: FEI VITROBOT MARK II / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 292 K
Details: 10nm colloidal gold was added to the sample prior to plunge freezing

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
MicroscopyModel: FEI TITAN KRIOS / Details: Nanoprobe
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM
Electron lensMode: BRIGHT FIELD / Nominal magnification: 81000 X / Nominal defocus max: 6500 nm / Nominal defocus min: 2000 nm / Cs: 2.7 mm / C2 aperture diameter: 50 µm / Alignment procedure: ZEMLIN TABLEAU
Specimen holderCryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER
Image recordingElectron dose: 2.2 e/Å2 / Detector mode: SUPER-RESOLUTION / Film or detector model: GATAN K2 QUANTUM (4k x 4k) / Num. of grids imaged: 1
EM imaging opticsEnergyfilter name: GIF Quantum LS / Energyfilter upper: 20 eV / Energyfilter lower: 0 eV
Image scansWidth: 3710 / Height: 3838 / Movie frames/image: 5

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Processing

EM software
IDNameCategory
2SerialEMimage acquisition
4IMODCTF correction
7UCSF Chimeramodel fitting
12AV3final Euler assignment
13TOM Toolboxfinal Euler assignment
15AV33D reconstruction
16TOM Toolbox3D reconstruction
Image processingDetails: Frames were aligned using MotionCorr. Tilts in a tilt series were exposure filtered for cumulative electron dose. Tomograms were reconstructed using IMOD.
CTF correctionDetails: CTF correction was performed using the ctfphaseflip program in IMOD prior to backprojection.
Type: PHASE FLIPPING ONLY
3D reconstructionResolution: 8.5 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 9161
Details: The final reconstruction is obtained by averaging, without further alignment, subtomograms that were selected based on the local curvature of the hexameric lattice. This reconstruction has ...Details: The final reconstruction is obtained by averaging, without further alignment, subtomograms that were selected based on the local curvature of the hexameric lattice. This reconstruction has hexamer-hexamer curvature parameters: tilt=11, twist=12.
Num. of class averages: 1 / Symmetry type: POINT
EM volume selectionDetails: Subtomograms were extracted along the manually rendered core surface of each viral particle.
Num. of tomograms: 103 / Num. of volumes extracted: 652618 / Reference model: reference free
Atomic model buildingProtocol: RIGID BODY FIT / Space: REAL / Target criteria: Cross-correlation coefficient

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