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Yorodumi- PDB-8kgn: Structure of African swine fever virus topoisomerase II in comple... -
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-Basic information
Entry | Database: PDB / ID: 8kgn | ||||||
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Title | Structure of African swine fever virus topoisomerase II in complex with dsDNA | ||||||
Components |
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Keywords | VIRAL PROTEIN/DNA / topo 2 / VIRAL PROTEIN-DNA complex | ||||||
Function / homology | Function and homology information sister chromatid segregation / DNA topoisomerase type II (double strand cut, ATP-hydrolyzing) activity / DNA topoisomerase (ATP-hydrolysing) / DNA topological change / DNA binding / ATP binding / metal ion binding / cytoplasm Similarity search - Function | ||||||
Biological species | African swine fever virus DNA molecule (others) | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 5.9 Å | ||||||
Authors | Cong, J. / Xin, Y. / Li, X. / Chen, Y. | ||||||
Funding support | 1items
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Citation | Journal: Nat Commun / Year: 2024 Title: Structural insights into the DNA topoisomerase II of the African swine fever virus. Authors: Jingyuan Cong / Yuhui Xin / Huiling Kang / Yunge Yang / Chenlong Wang / Dongming Zhao / Xuemei Li / Zihe Rao / Yutao Chen / Abstract: Type II topoisomerases are ubiquitous enzymes that play a pivotal role in modulating the topological configuration of double-stranded DNA. These topoisomerases are required for DNA metabolism and ...Type II topoisomerases are ubiquitous enzymes that play a pivotal role in modulating the topological configuration of double-stranded DNA. These topoisomerases are required for DNA metabolism and have been extensively studied in both prokaryotic and eukaryotic organisms. However, our understanding of virus-encoded type II topoisomerases remains limited. One intriguing example is the African swine fever virus, which stands as the sole mammalian-infecting virus encoding a type II topoisomerase. In this work, we use several approaches including cryo-EM, X-ray crystallography, and biochemical assays to investigate the structure and function of the African swine fever virus type II topoisomerase, pP1192R. We determine the structures of pP1192R in different conformational states and confirm its enzymatic activity in vitro. Collectively, our results illustrate the basic mechanisms of viral type II topoisomerases, increasing our understanding of these enzymes and presenting a potential avenue for intervention strategies to mitigate the impact of the African swine fever virus. | ||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 8kgn.cif.gz | 630.1 KB | Display | PDBx/mmCIF format |
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PDB format | pdb8kgn.ent.gz | 509.8 KB | Display | PDB format |
PDBx/mmJSON format | 8kgn.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 8kgn_validation.pdf.gz | 1.3 MB | Display | wwPDB validaton report |
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Full document | 8kgn_full_validation.pdf.gz | 1.3 MB | Display | |
Data in XML | 8kgn_validation.xml.gz | 81.9 KB | Display | |
Data in CIF | 8kgn_validation.cif.gz | 122.2 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/kg/8kgn ftp://data.pdbj.org/pub/pdb/validation_reports/kg/8kgn | HTTPS FTP |
-Related structure data
Related structure data | 37227MC 8kglC 8kgmC 8kgoC 8kgpC 8kgqC 8kgrC 8kgsC 8kgtC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
#1: Protein | Mass: 138093.359 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) African swine fever virus / Gene: P1192R CDS / Production host: Spodoptera frugiperda (fall armyworm) / References: UniProt: A0A2X0THW2 #2: DNA chain | | Mass: 16058.391 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) DNA molecule (others) #3: DNA chain | | Mass: 15965.361 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) DNA molecule (others) |
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-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 | Name: pP1192R / Type: COMPLEX / Details: dimer / Entity ID: #1 / Source: RECOMBINANT |
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Source (natural) | Organism: African swine fever virus |
Source (recombinant) | Organism: Spodoptera frugiperda (fall armyworm) |
Buffer solution | pH: 7.2 |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 2500 nm / Nominal defocus min: 1200 nm |
Image recording | Electron dose: 60 e/Å2 / Film or detector model: GATAN K2 SUMMIT (4k x 4k) |
-Processing
CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
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3D reconstruction | Resolution: 5.9 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 15000 / Symmetry type: POINT | ||||||||||||||||||||||||
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