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- PDB-3ja5: Genome and RdRp structure within the capsid of no-transcribing cy... -

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Entry
Database: PDB / ID: 3ja5
TitleGenome and RdRp structure within the capsid of no-transcribing cypovirus
ComponentsRNA-dependent RNA polymerase
KeywordsTRANSFERASE / non-transcribing cypovirus / RNA-dependent RNA polymerase / dsRNA virus / icosahedral virus
Function / homologyRNA-directed RNA polymerase, reovirus / RdRp of Reoviridae dsRNA viruses catalytic domain profile. / viral genome replication / RNA-dependent RNA polymerase activity / RNA binding / RNA-dependent RNA polymerase
Function and homology information
Biological speciesBombyx mori cypovirus 1
MethodELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 12 Å
AuthorsLiu, H. / Cheng, L.
CitationJournal: Science / Year: 2015
Title: Cryo-EM shows the polymerase structures and a nonspooled genome within a dsRNA virus.
Authors: Hongrong Liu / Lingpeng Cheng /
Abstract: Double-stranded RNA (dsRNA) viruses possess a segmented dsRNA genome and a number of RNA-dependent RNA polymerases (RdRps) enclosed in a capsid. Until now, the precise structures of genomes and RdRps ...Double-stranded RNA (dsRNA) viruses possess a segmented dsRNA genome and a number of RNA-dependent RNA polymerases (RdRps) enclosed in a capsid. Until now, the precise structures of genomes and RdRps within the capsids have been unknown. Here we report the structures of RdRps and associated RNAs within nontranscribing and transcribing cypoviruses (NCPV and TCPV, respectively), using a combination of cryo-electron microscopy (cryo-EM) and a symmetry-mismatch reconstruction method. The RdRps and associated RNAs appear to exhibit a pseudo-D3 symmetric organization in both NCPV and TCPV. However, the molecular interactions between RdRps and the genomic RNA were found to differ in these states. Our work provides insight into the mechanisms of the replication and transcription in dsRNA viruses and paves a way for structural determination of lower-symmetry complexes enclosed in higher-symmetry structures.
History
DepositionApr 21, 2015Deposition site: RCSB / Processing site: PDBJ
Revision 1.0Oct 28, 2015Provider: repository / Type: Initial release
Revision 1.1Jun 20, 2018Group: Data collection / Database references ...Data collection / Database references / Derived calculations / Other
Category: cell / database_2 ...cell / database_2 / em_image_scans / pdbx_struct_assembly / pdbx_struct_assembly_gen / pdbx_struct_oper_list
Item: _cell.length_a / _cell.length_b / _cell.length_c
Revision 1.2Dec 18, 2019Group: Data collection / Category: em_software / Item: _em_software.image_processing_id
Revision 1.3Mar 20, 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: RNA-dependent RNA polymerase


Theoretical massNumber of molelcules
Total (without water)138,8301
Polymers138,8301
Non-polymers00
Water0
1


  • Idetical with deposited unit
  • defined by author
TypeNameSymmetry operationNumber
identity operation1_5551
SymmetryPoint symmetry: (Schoenflies symbol: D3 (2x3 fold dihedral))

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Components

#1: Protein RNA-dependent RNA polymerase / / Coordinate model: Cα atoms only


Mass: 138830.109 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Bombyx mori cypovirus 1 / References: UniProt: Q6RJR5

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Experimental details

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Experiment

ExperimentMethod: ELECTRON MICROSCOPY
EM experimentAggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction

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Sample preparation

ComponentName: Bombyx mori cypovirus 1 / Type: VIRUS
Details of virusEmpty: NO / Enveloped: NO / Host category: VERTEBRATES / Isolate: SPECIES / Type: VIRION
Natural hostOrganism: Homo sapiens
SpecimenEmbedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
VitrificationInstrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
MicroscopyModel: FEI TITAN KRIOS / Date: Apr 10, 2010
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: OTHER
Electron lensMode: BRIGHT FIELDBright-field microscopy / Nominal magnification: 75000 X / Calibrated magnification: 125390 X / Camera length: 0 mm
Specimen holderSpecimen holder model: GATAN HELIUM / Tilt angle max: 0 ° / Tilt angle min: 0 °
Image recordingFilm or detector model: GATAN ULTRASCAN 4000 (4k x 4k)

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Processing

EM softwareName: MRC / Category: 3D reconstruction / Details: Symmetry-Mismatch-Reconstruction
CTF correctionDetails: Each particle
SymmetryPoint symmetry: D3 (2x3 fold dihedral)
3D reconstructionMethod: cross-correlation coefficient / Resolution: 12 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 28000
Details: (Single particle details: Symmetry-mismatch reconstruction of icosahedral with D3 symmetry imposed) (Single particle--Applied symmetry: D3)
Symmetry type: POINT
Refinement stepCycle: LAST
ProteinNucleic acidLigandSolventTotal
Num. atoms1074 0 0 0 1074

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