+Open data
-Basic information
Entry | Database: PDB / ID: 8hf2 | |||||||||
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Title | Cryo-EM structure of WeiTsing | |||||||||
Components | PRA1 family protein | |||||||||
Keywords | MEMBRANE PROTEIN / Pentamer / Channel | |||||||||
Function / homology | membrane / Transmembrane protein Function and homology information | |||||||||
Biological species | Arabidopsis thaliana (thale cress) | |||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 4.14 Å | |||||||||
Authors | Qin, L. / Tang, L.H. / Chen, Y.H. | |||||||||
Funding support | China, 2items
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Citation | Journal: Cell / Year: 2023 Title: WeiTsing, a pericycle-expressed ion channel, safeguards the stele to confer clubroot resistance. Authors: Wei Wang / Li Qin / Wenjing Zhang / Linghui Tang / Chao Zhang / Xiaojing Dong / Pei Miao / Meng Shen / Huilong Du / Hangyuan Cheng / Ke Wang / Xiangyun Zhang / Min Su / Hongwei Lu / Chang Li ...Authors: Wei Wang / Li Qin / Wenjing Zhang / Linghui Tang / Chao Zhang / Xiaojing Dong / Pei Miao / Meng Shen / Huilong Du / Hangyuan Cheng / Ke Wang / Xiangyun Zhang / Min Su / Hongwei Lu / Chang Li / Qiang Gao / Xiaojuan Zhang / Yun Huang / Chengzhi Liang / Jian-Min Zhou / Yu-Hang Chen / Abstract: Plant roots encounter numerous pathogenic microbes that often cause devastating diseases. One such pathogen, Plasmodiophora brassicae (Pb), causes clubroot disease and severe yield losses on ...Plant roots encounter numerous pathogenic microbes that often cause devastating diseases. One such pathogen, Plasmodiophora brassicae (Pb), causes clubroot disease and severe yield losses on cruciferous crops worldwide. Here, we report the isolation and characterization of WeiTsing (WTS), a broad-spectrum clubroot resistance gene from Arabidopsis. WTS is transcriptionally activated in the pericycle upon Pb infection to prevent pathogen colonization in the stele. Brassica napus carrying the WTS transgene displayed strong resistance to Pb. WTS encodes a small protein localized in the endoplasmic reticulum (ER), and its expression in plants induces immune responses. The cryoelectron microscopy (cryo-EM) structure of WTS revealed a previously unknown pentameric architecture with a central pore. Electrophysiology analyses demonstrated that WTS is a calcium-permeable cation-selective channel. Structure-guided mutagenesis indicated that channel activity is strictly required for triggering defenses. The findings uncover an ion channel analogous to resistosomes that triggers immune signaling in the pericycle. | |||||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 8hf2.cif.gz | 109.5 KB | Display | PDBx/mmCIF format |
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PDB format | pdb8hf2.ent.gz | 85.5 KB | Display | PDB format |
PDBx/mmJSON format | 8hf2.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 8hf2_validation.pdf.gz | 1 MB | Display | wwPDB validaton report |
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Full document | 8hf2_full_validation.pdf.gz | 1.1 MB | Display | |
Data in XML | 8hf2_validation.xml.gz | 26.2 KB | Display | |
Data in CIF | 8hf2_validation.cif.gz | 37.9 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/hf/8hf2 ftp://data.pdbj.org/pub/pdb/validation_reports/hf/8hf2 | HTTPS FTP |
-Related structure data
Related structure data | 34710MC 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: 16003.990 Da / Num. of mol.: 5 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Arabidopsis thaliana (thale cress) / Gene: AN1_LOCUS3247, C24_LOCUS3162 / Production host: Schizosaccharomyces pombe (fission yeast) / References: UniProt: A0A654EJS8 |
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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: WeiTsing / Type: COMPLEX / Entity ID: all / Source: RECOMBINANT |
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Source (natural) | Organism: Arabidopsis thaliana (thale cress) |
Source (recombinant) | Organism: Schizosaccharomyces pombe (fission yeast) |
Buffer solution | pH: 8 |
Specimen | Conc.: 2.5 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Specimen support | Grid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R1.2/1.3 |
Vitrification | Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277 K |
-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: 2000 nm / Nominal defocus min: 1200 nm / Cs: 2.7 mm |
Image recording | Electron dose: 50 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
-Processing
Software | Name: PHENIX / Version: dev_3689: / Classification: refinement | ||||||||||||||||||||||||
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EM software |
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CTF correction | Type: NONE | ||||||||||||||||||||||||
Symmetry | Point symmetry: C5 (5 fold cyclic) | ||||||||||||||||||||||||
3D reconstruction | Resolution: 4.14 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 117360 / Symmetry type: POINT | ||||||||||||||||||||||||
Refine LS restraints |
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