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- EMDB-51810: Cryo-EM structure of an octameric G10-resistosome from wheat -

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Basic information

Entry
Database: EMDB / ID: EMD-51810
TitleCryo-EM structure of an octameric G10-resistosome from wheat
Map datapostprocess map used for model building
Sample
  • Complex: Wheat auto immunity -3 Protein
    • Protein or peptide: NB-ARC domain-containing protein
  • Ligand: ADENOSINE-5'-TRIPHOSPHATE
KeywordsG10-NLR / Octamer / Resistance / LRR / PLANT PROTEIN
Function / homology
Function and homology information


plant-type hypersensitive response / innate immune response-activating signaling pathway / ADP binding / defense response to bacterium / ATP binding
Similarity search - Function
: / : / Plant disease resistance protein RPS2-like, leucine-rich repeats / : / Disease resistance protein Winged helix domain / Apoptotic protease-activating factors, helical domain / NB-ARC / NB-ARC domain / Leucine rich repeat / Leucine-rich repeat, typical subtype ...: / : / Plant disease resistance protein RPS2-like, leucine-rich repeats / : / Disease resistance protein Winged helix domain / Apoptotic protease-activating factors, helical domain / NB-ARC / NB-ARC domain / Leucine rich repeat / Leucine-rich repeat, typical subtype / Leucine-rich repeats, typical (most populated) subfamily / Leucine-rich repeat profile. / Leucine-rich repeat / Leucine-rich repeat domain superfamily / Winged helix-like DNA-binding domain superfamily / P-loop containing nucleoside triphosphate hydrolase
Similarity search - Domain/homology
Uncharacterized protein
Similarity search - Component
Biological speciesTriticum aestivum (bread wheat)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.95 Å
AuthorsGuo GH / Zhao H / Lukoyanova N / Selvaraj M / Jones J
Funding support United Kingdom, 1 items
OrganizationGrant numberCountry
Gatsby Charitable Foundation United Kingdom
Citation
Journal: Cell / Year: 2026
Title: An activated wheat CC-NLR immune receptor forms an octameric resistosome.
Authors: Guanghao Guo / He Zhao / Kaihong Bai / Jian Lu / Qiuhong Wu / Lei Lu / Yue Zhang / Lingli Dong / Guangwei Li / Yongxing Chen / Yikun Hou / Ping Lu / Miaomiao Li / Huaizhi Zhang / Gaojie Wang ...Authors: Guanghao Guo / He Zhao / Kaihong Bai / Jian Lu / Qiuhong Wu / Lei Lu / Yue Zhang / Lingli Dong / Guangwei Li / Yongxing Chen / Yikun Hou / Ping Lu / Miaomiao Li / Huaizhi Zhang / Gaojie Wang / Keyu Zhu / Baoge Huang / Xuejia Cui / Hongkui Fu / Chenchen Hu / Zhiying Chu / Xue Lyu / Sophien Kamoun / Chao Wang / Zhiyong Liu / Muniyandi Selvaraj / Jonathan D G Jones /
Abstract: Nucleotide-binding, leucine-rich repeat (NLR) receptors are widespread intracellular immune sensors across kingdoms. Plant G10-type coiled-coil (CC)-NLRs constitute a distinct phylogenetic clade that ...Nucleotide-binding, leucine-rich repeat (NLR) receptors are widespread intracellular immune sensors across kingdoms. Plant G10-type coiled-coil (CC)-NLRs constitute a distinct phylogenetic clade that remains poorly characterized. Here, we identified a gain-of-function mutant of wheat autoimmunity 3 (WAI3), which encodes a constitutively active CC-NLR resulting from a residue substitution in the leucine-rich repeat (LRR) domain. Cryo-electron microscopy (cryo-EM) analysis reveals that activated WAI3 assembles into a distinctive octameric resistosome. Arabidopsis RPS2, another CC-NLR, also forms an octamer, indicating a conserved structural property across monocot and dicot plants. The WAI3 resistosome induces a prolonged and sustained increase in cytosolic calcium, likely facilitated by a unique channel architecture arising from its divergent coiled-coil (CC) domain configuration. Notably, this domain arrangement may be shared by plant NLRs that lack the conserved EDVID (Glu-Asp-Val-Ile-Asp) motif in their CC domains. Together, our findings uncover a conserved yet previously uncharacterized NLR resistosome structure and provide insights into the plant immune receptor plasticity.
#1: Journal: Biorxiv / Year: 2025
Title: An activated wheat CCG10-NLR immune receptor forms an octameric resistosome
Authors: Guo G / Zhao H / Bai K / Wu Q / Dong L / Lu L / Chen Y / Hou Y / Lu J / Lu P / Li M / Zhang H / Wang G / Zhu K / Huang B / Cui X / Fu H / Hu C / Chu Z / Lyu X / Kamoun S / Wang C / Liu Z / Selvaraj M / Jones JDG
History
DepositionOct 11, 2024-
Header (metadata) releaseSep 24, 2025-
Map releaseSep 24, 2025-
UpdateApr 1, 2026-
Current statusApr 1, 2026Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_51810.png

Downloads & links

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Map

FileDownload / File: emd_51810.map.gz / Format: CCP4 / Size: 512 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotationpostprocess map used for model building
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.83 Å/pix.
x 512 pix.
= 422.912 Å		0.83 Å/pix.
x 512 pix.
= 422.912 Å		0.83 Å/pix.
x 512 pix.
= 422.912 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 0.826 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.008
Minimum - Maximum-0.02534979 - 0.05313489
Average (Standard dev.)0.0002070895 (±0.0015597356)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions512512512
Spacing512512512
CellA=B=C: 422.912 Å
α=β=γ: 90.0 °

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Supplemental data

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Mask #1

Fileemd_51810_msk_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: Thisis half map 1

Fileemd_51810_half_map_1.map
AnnotationThisis half map 1
Projections & Slices
AxesZYX

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Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: This is half map 2

Fileemd_51810_half_map_2.map
AnnotationThis is half map 2
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : Wheat auto immunity -3 Protein

EntireName: Wheat auto immunity -3 Protein
Components
  • Complex: Wheat auto immunity -3 Protein
    • Protein or peptide: NB-ARC domain-containing protein
  • Ligand: ADENOSINE-5'-TRIPHOSPHATE

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Supramolecule #1: Wheat auto immunity -3 Protein

SupramoleculeName: Wheat auto immunity -3 Protein / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1 / Details: This is a resistance protein in the wheat plant
Source (natural)Organism: Triticum aestivum (bread wheat) / Organ: Leaves / Tissue: leaf
Molecular weightTheoretical: 856 KDa

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Macromolecule #1: NB-ARC domain-containing protein

MacromoleculeName: NB-ARC domain-containing protein / type: protein_or_peptide / ID: 1 / Details: This is a wheat resistance protein in G10-clade. / Number of copies: 8 / Enantiomer: LEVO
Source (natural)Organism: Triticum aestivum (bread wheat) / Strain: M3405
Molecular weightTheoretical: 107.700422 KDa
Recombinant expressionOrganism: Nicotiana benthamiana (plant)
SequenceString: MADAISAAGS CEQPECECLD GTGMLDAAAR EVASFLHLKS NWSDLDKAKK LLLAVETTVR ARVTAEVDKL NICDPQVQVW LRRVEELQL DAIDEDYSQL RKYSCLGQCT IHAHRRASIG RRVLEALDEA NKLIEEGRRF KKFGFKPLPK IVDPLPQIKT F GLETMLSQ ...String:
MADAISAAGS CEQPECECLD GTGMLDAAAR EVASFLHLKS NWSDLDKAKK LLLAVETTVR ARVTAEVDKL NICDPQVQVW LRRVEELQL DAIDEDYSQL RKYSCLGQCT IHAHRRASIG RRVLEALDEA NKLIEEGRRF KKFGFKPLPK IVDPLPQIKT F GLETMLSQ LYDLFEKGDS NIIGVWGQGG VGKTTLLHVF NNDLEKKAHD YQVVIFIEVS NSEALNTVEI QQTISERLNL PW NDAEPIA KRARFLIKAL GRKRFVILLD DVRKKFCLED VGIPTPDINS QSKLILTSRY REVCFQMNAQ RSLIEMQILG NDA SWELFL SKLSTETSAA VEPLGSQSAT REHAMKIAQS CGGLPLALNV IGTAVAGLEE GEWQSAADAI ATNMDNIDGV DEMF GRLKY SFDRLTPTQQ QCFLYCTLFP EYGSISKEQL IGYWLAEGLL LNDSEKGYQI IRSLVSACLL QVSGSMSSKV KMHHV IRQL GLWLVNKSDT KFLVQPGMAL DNAPSAEEWN EATRISIMSN NITELSFSPK CKNVTTLLMQ NNPNLNKMSY GFFRTM SSL KVLDLSHTAI TSLPECDALV ALEHLNLSHT HIMRLPERLW LLKELRHLDL SVTVAFEDTM NNCSKLHKLK VLNLFRS HY GIRDVDNLNL DSLKELLFLG ITIYAEDVLK KLNMPRPLAK STHRLNLKYC AEMQSIKISD LSHMEHLEEL YVESCYDL N TVVADAELTT SQLQFLTLSV LPSLESVLVA PMSHNFQYIR KLIISHCPKL SNITWVRRLQ LLERLVISHC DGVLEIVED EEQYGEQMKM QDHASYEQEE HAMVETSRND TGQSDFPKLR LIVLTGLKKL RSICKAREFP CLETLRVEDC PNLRSIPLSC THNYWKLKQ ICGSVEWWEK LQWENRKEVA CLDSKYFIPI GSDYKDHDGD YKDHDLDAAA ADYKDDDDK

UniProtKB: Uncharacterized protein

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Macromolecule #2: ADENOSINE-5'-TRIPHOSPHATE

MacromoleculeName: ADENOSINE-5'-TRIPHOSPHATE / type: ligand / ID: 2 / Number of copies: 8 / Formula: ATP
Molecular weightTheoretical: 507.181 Da
Chemical component information

ChemComp-ATP:
ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying molecule*YM

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

BufferpH: 7.5 / Component - Concentration: 150.0 mM / Component - Formula: NaCl / Component - Name: Sodium chloride
Details: 150mM Tris HCl pH 7.5 150mM NaCl 1mM EDTA 1mM MgCl2
GridModel: Quantifoil R1.2/1.3 / Material: COPPER / Support film - Material: GRAPHENE / Pretreatment - Type: GLOW DISCHARGE
VitrificationCryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV
Details: 3 microlitre of the sample was applied on a graphene coated Quantafoil grid R1.2/1.3 copper grid and vitrification was performed using vitrobot.
Detailsthis is a protein sample purified from plant cell extract after over expression in leaf tissues

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Electron microscopy

MicroscopeTFS KRIOS
Specialist opticsEnergy filter - Name: GIF Bioquantum / Energy filter - Slit width: 10 eV
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 1 / Number real images: 7044 / Average exposure time: 1.94 sec. / Average electron dose: 50.0 e/Å2
Details: One grid was imaged at areas with good ice and several images
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.5 µm / Nominal defocus min: 1.3 µm / Nominal magnification: 105000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

CTF correctionSoftware - Name: RELION (ver. 4.0) / Type: PHASE FLIPPING ONLY
Startup modelType of model: OTHER / Details: ab-initio model reconstruction in relion
Final reconstructionApplied symmetry - Point group: C8 (8 fold cyclic) / Algorithm: BACK PROJECTION / Resolution.type: BY AUTHOR / Resolution: 3.95 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 5.0)
Details: The half-maps and resolution were obtained using RELION software
Number images used: 12203
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 5.0)
Final angle assignmentType: MAXIMUM LIKELIHOOD
FSC plot (resolution estimation)

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Atomic model buiding 1

Initial modelChain - Source name: AlphaFold / Chain - Initial model type: in silico model
DetailsThe atomic model was fitted flexibly using coot and refined using Phenix realspace refinement module
RefinementSpace: REAL / Protocol: FLEXIBLE FIT
Output model

PDB-9h2l:
Cryo-EM structure of an octameric G10-resistosome from wheat

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