+Open data
-Basic information
Entry | Database: PDB / ID: 7una | ||||||||||||||||||||||||||||||||||||
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Title | SfSTING with cGAMP (masked) | ||||||||||||||||||||||||||||||||||||
Components | CD-NTase-associated protein 12 | ||||||||||||||||||||||||||||||||||||
Keywords | ANTIVIRAL PROTEIN / STING / bacterial / filament | ||||||||||||||||||||||||||||||||||||
Function / homology | Function and homology information NAD+ glycohydrolase / NADP+ nucleosidase activity / NAD+ nucleosidase activity / defense response to virus / nucleotide binding Similarity search - Function | ||||||||||||||||||||||||||||||||||||
Biological species | Sphingobacterium faecium (bacteria) | ||||||||||||||||||||||||||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 4 Å | ||||||||||||||||||||||||||||||||||||
Authors | Morehouse, B.R. / Yip, M.C.J. / Keszei, A.F.A. / McNamara-Bordewick, N.K. / Shao, S. / Kranzusch, P.J. | ||||||||||||||||||||||||||||||||||||
Funding support | United States, 11items
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Citation | Journal: Nature / Year: 2022 Title: Cryo-EM structure of an active bacterial TIR-STING filament complex. Authors: Benjamin R Morehouse / Matthew C J Yip / Alexander F A Keszei / Nora K McNamara-Bordewick / Sichen Shao / Philip J Kranzusch / Abstract: Stimulator of interferon genes (STING) is an antiviral signalling protein that is broadly conserved in both innate immunity in animals and phage defence in prokaryotes. Activation of STING requires ...Stimulator of interferon genes (STING) is an antiviral signalling protein that is broadly conserved in both innate immunity in animals and phage defence in prokaryotes. Activation of STING requires its assembly into an oligomeric filament structure through binding of a cyclic dinucleotide, but the molecular basis of STING filament assembly and extension remains unknown. Here we use cryogenic electron microscopy to determine the structure of the active Toll/interleukin-1 receptor (TIR)-STING filament complex from a Sphingobacterium faecium cyclic-oligonucleotide-based antiphage signalling system (CBASS) defence operon. Bacterial TIR-STING filament formation is driven by STING interfaces that become exposed on high-affinity recognition of the cognate cyclic dinucleotide signal c-di-GMP. Repeating dimeric STING units stack laterally head-to-head through surface interfaces, which are also essential for human STING tetramer formation and downstream immune signalling in mammals. The active bacterial TIR-STING structure reveals further cross-filament contacts that brace the assembly and coordinate packing of the associated TIR NADase effector domains at the base of the filament to drive NAD hydrolysis. STING interface and cross-filament contacts are essential for cell growth arrest in vivo and reveal a stepwise mechanism of activation whereby STING filament assembly is required for subsequent effector activation. Our results define the structural basis of STING filament formation in prokaryotic antiviral signalling. | ||||||||||||||||||||||||||||||||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 7una.cif.gz | 404 KB | Display | PDBx/mmCIF format |
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PDB format | pdb7una.ent.gz | 327.7 KB | Display | PDB format |
PDBx/mmJSON format | 7una.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 7una_validation.pdf.gz | 1.4 MB | Display | wwPDB validaton report |
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Full document | 7una_full_validation.pdf.gz | 1.4 MB | Display | |
Data in XML | 7una_validation.xml.gz | 51.7 KB | Display | |
Data in CIF | 7una_validation.cif.gz | 72.1 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/un/7una ftp://data.pdbj.org/pub/pdb/validation_reports/un/7una | HTTPS FTP |
-Related structure data
Related structure data | 26618MC 7un8C 7un9C C: citing same article (ref.) M: map data used to model this data |
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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: 36956.051 Da / Num. of mol.: 8 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Sphingobacterium faecium (bacteria) / Gene: cap12, C8N37_104320, SF1_08920 / Production host: Escherichia coli (E. coli) / References: UniProt: A0A2T5Y4G4, NAD+ glycohydrolase #2: Chemical | ChemComp-4BW / Has ligand of interest | Y | |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: FILAMENT / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: SfSTING with cGAMP / Type: COMPLEX / Entity ID: #1 / Source: RECOMBINANT |
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Molecular weight | Experimental value: NO |
Source (natural) | Organism: Sphingobacterium faecium (bacteria) |
Source (recombinant) | Organism: Escherichia coli (E. coli) |
Buffer solution | pH: 7.5 |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy imaging
Experimental equipment | Model: Talos Arctica / Image courtesy: FEI Company |
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Microscopy | Model: FEI TALOS ARCTICA |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 200 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 2600 nm / Nominal defocus min: 1400 nm |
Image recording | Electron dose: 52.9 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
-Processing
CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION |
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3D reconstruction | Resolution: 4 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 105567 / Symmetry type: POINT |