+Open data
-Basic information
Entry | Database: PDB / ID: 5h0r | ||||||||||||||||||
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Title | RNA dependent RNA polymerase ,vp4,dsRNA | ||||||||||||||||||
Components |
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Keywords | TRANSFERASE/RNA / structural classification / TRANSFERASE-RNA complex | ||||||||||||||||||
Function / homology | Function and homology information viral genome replication / RNA-dependent RNA polymerase activity / RNA binding Similarity search - Function | ||||||||||||||||||
Biological species | Bombyx mori cytoplasmic polyhedrosis virus Dendrolimus punctatus cypovirus 1 Cypovirus (cytoplasmic polyhedrosis viruses) | ||||||||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.9 Å | ||||||||||||||||||
Authors | Li, X. / Zhou, N. / Chen, W. / Zhu, B. / Wang, X. / Xu, B. / Wang, J. / Liu, H. / Cheng, L. | ||||||||||||||||||
Funding support | China, 5items
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Citation | Journal: J Mol Biol / Year: 2017 Title: Near-Atomic Resolution Structure Determination of a Cypovirus Capsid and Polymerase Complex Using Cryo-EM at 200kV. Authors: Xiaowu Li / Niyun Zhou / Wenyuan Chen / Bin Zhu / Xurong Wang / Bin Xu / Jiawei Wang / Hongrong Liu / Lingpeng Cheng / Abstract: Single-particle cryo-electron microscopy (cryo-EM) allows the high-resolution structural determination of biological assemblies in a near-native environment. However, all high-resolution (better than ...Single-particle cryo-electron microscopy (cryo-EM) allows the high-resolution structural determination of biological assemblies in a near-native environment. However, all high-resolution (better than 3.5Å) cryo-EM structures reported to date were obtained by using 300kV transmission electron microscopes (TEMs). We report here the structures of a cypovirus capsid of 750-Å diameter at 3.3-Å resolution and of RNA-dependent RNA polymerase (RdRp) complexes within the capsid at 3.9-Å resolution using a 200-kV TEM. The newly resolved structure revealed conformational changes of two subdomains in the RdRp. These conformational changes, which were involved in RdRp's switch from non-transcribing to transcribing mode, suggest that the RdRp may facilitate the unwinding of genomic double-stranded RNA. The possibility of 3-Å resolution structural determinations for biological assemblies of relatively small sizes using cryo-EM at 200kV was discussed. | ||||||||||||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | Molecule: MolmilJmol/JSmol |
-Downloads & links
-Download
PDBx/mmCIF format | 5h0r.cif.gz | 371.4 KB | Display | PDBx/mmCIF format |
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PDB format | pdb5h0r.ent.gz | 290.4 KB | Display | PDB format |
PDBx/mmJSON format | 5h0r.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 5h0r_validation.pdf.gz | 1.3 MB | Display | wwPDB validaton report |
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Full document | 5h0r_full_validation.pdf.gz | 1.3 MB | Display | |
Data in XML | 5h0r_validation.xml.gz | 58.5 KB | Display | |
Data in CIF | 5h0r_validation.cif.gz | 87.4 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/h0/5h0r ftp://data.pdbj.org/pub/pdb/validation_reports/h0/5h0r | HTTPS FTP |
-Related structure data
Related structure data | 9564MC 9565C 5h0sC C: citing same article (ref.) M: map data used to model this data |
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Similar structure data |
-Links
-Assembly
Deposited unit |
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1 |
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2 |
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-Components
#1: Protein | Mass: 139077.281 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Bombyx mori cytoplasmic polyhedrosis virus Gene: RdRp Production host: Cypovirus (cytoplasmic polyhedrosis viruses) References: UniProt: A0A0S1LIW6 |
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#2: Protein | Mass: 63734.707 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Dendrolimus punctatus cypovirus 1 Production host: Cypovirus (cytoplasmic polyhedrosis viruses) References: UniProt: Q80A92 |
#3: RNA chain | Mass: 13781.683 Da / Num. of mol.: 1 / Source method: obtained synthetically Source: (synth.) Cypovirus (cytoplasmic polyhedrosis viruses) |
#4: RNA chain | Mass: 12814.002 Da / Num. of mol.: 1 / Source method: obtained synthetically Source: (synth.) Cypovirus (cytoplasmic polyhedrosis viruses) |
-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component |
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Source (recombinant) |
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Buffer solution | pH: 7.5 | ||||||||||||||||||||||||||||||||||||
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | ||||||||||||||||||||||||||||||||||||
Vitrification | Cryogen name: NITROGEN |
-Electron microscopy imaging
Experimental equipment | Model: Talos Arctica / Image courtesy: FEI Company |
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Microscopy | Model: FEI TECNAI ARCTICA |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 200 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD |
Image recording | Electron dose: 20 e/Å2 / Film or detector model: FEI FALCON II (4k x 4k) |
-Processing
Software | Name: PHENIX / Version: 1.10.1_2155: / Classification: refinement | ||||||||||||||||||||||||
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
3D reconstruction | Resolution: 3.9 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 27000 / Symmetry type: POINT | ||||||||||||||||||||||||
Refine LS restraints |
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