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- PDB-7v9u: Cryo-EM structure of E.coli retron-Ec86 (RT-msDNA-RNA) at 3.2 angstrom -
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Open data
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Basic information
Entry | Database: PDB / ID: 7v9u | |||||||||
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Title | Cryo-EM structure of E.coli retron-Ec86 (RT-msDNA-RNA) at 3.2 angstrom | |||||||||
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![]() | TRANSFERASE/DNA/RNA / Reverse transcriptase / RNA BINDING PROTEIN / TRANSFERASE-DNA-RNA complex | |||||||||
Function / homology | ![]() RNA-directed DNA polymerase / RNA-directed DNA polymerase activity / defense response to virus / RNA binding / metal ion binding Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.12 Å | |||||||||
![]() | Wang, Y.J. / Guan, Z.Y. / Zou, T.T. | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Cryo-EM structures of Escherichia coli Ec86 retron complexes reveal architecture and defence mechanism. Authors: Yanjing Wang / Zeyuan Guan / Chen Wang / Yangfan Nie / Yibei Chen / Zhaoyang Qian / Yongqing Cui / Han Xu / Qiang Wang / Fen Zhao / Delin Zhang / Pan Tao / Ming Sun / Ping Yin / Shuangxia ...Authors: Yanjing Wang / Zeyuan Guan / Chen Wang / Yangfan Nie / Yibei Chen / Zhaoyang Qian / Yongqing Cui / Han Xu / Qiang Wang / Fen Zhao / Delin Zhang / Pan Tao / Ming Sun / Ping Yin / Shuangxia Jin / Shan Wu / Tingting Zou / ![]() Abstract: First discovered in the 1980s, retrons are bacterial genetic elements consisting of a reverse transcriptase and a non-coding RNA (ncRNA). Retrons mediate antiphage defence in bacteria but their ...First discovered in the 1980s, retrons are bacterial genetic elements consisting of a reverse transcriptase and a non-coding RNA (ncRNA). Retrons mediate antiphage defence in bacteria but their structure and defence mechanisms are unknown. Here, we investigate the Escherichia coli Ec86 retron and use cryo-electron microscopy to determine the structures of the Ec86 (3.1 Å) and cognate effector-bound Ec86 (2.5 Å) complexes. The Ec86 reverse transcriptase exhibits a characteristic right-hand-like fold consisting of finger, palm and thumb subdomains. Ec86 reverse transcriptase reverse-transcribes part of the ncRNA into satellite, multicopy single-stranded DNA (msDNA, a DNA-RNA hybrid) that we show wraps around the reverse transcriptase electropositive surface. In msDNA, both inverted repeats are present and the 3' sides of the DNA/RNA chains are close to the reverse transcriptase active site. The Ec86 effector adopts a two-lobe fold and directly binds reverse transcriptase and msDNA. These findings offer insights into the structure-function relationship of the retron-effector unit and provide a structural basis for the optimization of retron-based genome editing systems. | |||||||||
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Structure visualization
Structure viewer | Molecule: ![]() ![]() |
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PDBx/mmCIF format | ![]() | 220.5 KB | Display | ![]() |
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PDB format | ![]() | 166.5 KB | Display | ![]() |
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-Validation report
Summary document | ![]() | 866.8 KB | Display | ![]() |
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Full document | ![]() | 883 KB | Display | |
Data in XML | ![]() | 28 KB | Display | |
Data in CIF | ![]() | 43.1 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 31827MC ![]() 7xjgC M: map data used to model this data C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
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Assembly
Deposited unit | ![]()
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Components
#1: Protein | Mass: 36484.641 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() ![]() ![]() #2: DNA chain | Mass: 32468.803 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() ![]() ![]() #3: RNA chain | Mass: 25870.203 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() ![]() ![]() #4: RNA chain | Mass: 4541.771 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() ![]() ![]() |
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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 |
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Sample preparation
Component | Name: retron-Ec86 (RT-msDNA-RNA) / Type: COMPLEX / Entity ID: all / Source: RECOMBINANT |
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Source (natural) | Organism: ![]() ![]() |
Source (recombinant) | Organism: ![]() ![]() |
Buffer solution | pH: 8 |
Specimen | Conc.: 0.4 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Vitrification | Cryogen name: ETHANE |
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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: ![]() |
Electron lens | Mode: BRIGHT FIELD |
Image recording | Electron dose: 53.68 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
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Processing
Software | Name: PHENIX / Version: 1.19.1_4122: / Classification: refinement | ||||||||||||||||||||||||
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EM software | Name: cryoSPARC / Version: 3.1 / Category: final Euler assignment | ||||||||||||||||||||||||
CTF correction | Type: NONE | ||||||||||||||||||||||||
Symmetry | Point symmetry: C2 (2 fold cyclic) | ||||||||||||||||||||||||
3D reconstruction | Resolution: 3.12 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 144586 / Symmetry type: POINT | ||||||||||||||||||||||||
Refine LS restraints |
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