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Yorodumi- PDB-6tc3: Cryo-EM structure of an Escherichia coli ribosome-SpeFL complex s... -
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Basic information
| Entry | Database: PDB / ID: 6tc3 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Title | Cryo-EM structure of an Escherichia coli ribosome-SpeFL complex stalled in response to L-ornithine (Replicate 1) | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|  Components | 
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|  Keywords | RIBOSOME / SpeFL / arrest peptide / ornithine / protein synthesis | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Function / homology |  Function and homology information transcriptional attenuation by ribosome / regulation of translational elongation / negative regulation of cytoplasmic translational initiation / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / endoribonuclease inhibitor activity ...transcriptional attenuation by ribosome / regulation of translational elongation / negative regulation of cytoplasmic translational initiation / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / endoribonuclease inhibitor activity / positive regulation of ribosome biogenesis / RNA-binding transcription regulator activity / negative regulation of cytoplasmic translation / four-way junction DNA binding / DnaA-L2 complex / translation repressor activity / negative regulation of translational initiation / regulation of mRNA stability / negative regulation of DNA-templated DNA replication initiation / mRNA regulatory element binding translation repressor activity / assembly of large subunit precursor of preribosome / positive regulation of RNA splicing / ribosome assembly / regulation of DNA-templated transcription elongation / transcription elongation factor complex / cytosolic ribosome assembly / response to reactive oxygen species / DNA endonuclease activity / transcription antitermination / translational initiation / regulation of cell growth / DNA-templated transcription termination / response to radiation / maintenance of translational fidelity / mRNA 5'-UTR binding / ribosome biogenesis / large ribosomal subunit / regulation of translation / ribosome binding / transferase activity / ribosomal small subunit biogenesis / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / small ribosomal subunit rRNA binding / ribosomal large subunit assembly / cytosolic small ribosomal subunit / large ribosomal subunit rRNA binding / cytosolic large ribosomal subunit / cytoplasmic translation / tRNA binding / negative regulation of translation / rRNA binding / structural constituent of ribosome / ribosome / translation / response to antibiotic / negative regulation of DNA-templated transcription / mRNA binding / DNA binding / RNA binding / zinc ion binding / membrane / cytoplasm / cytosol Similarity search - Function | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Biological species |   Escherichia coli K-12 (bacteria) | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.7 Å | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|  Authors | Innis, C.A. / Herrero del Valle, A. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Funding support | 3items 
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|  Citation |  Journal: Nat Microbiol / Year: 2020 Title: Ornithine capture by a translating ribosome controls bacterial polyamine synthesis. Authors: Alba Herrero Del Valle / Britta Seip / Iñaki Cervera-Marzal / Guénaël Sacheau / A Carolin Seefeldt / C Axel Innis /      Abstract: Polyamines are essential metabolites that play an important role in cell growth, stress adaptation and microbial virulence. To survive and multiply within a human host, pathogenic bacteria adjust the ...Polyamines are essential metabolites that play an important role in cell growth, stress adaptation and microbial virulence. To survive and multiply within a human host, pathogenic bacteria adjust the expression and activity of polyamine biosynthetic enzymes in response to different environmental stresses and metabolic cues. Here, we show that ornithine capture by the ribosome and the nascent peptide SpeFL controls polyamine synthesis in γ-proteobacteria by inducing the expression of the ornithine decarboxylase SpeF, via a mechanism involving ribosome stalling and transcription antitermination. In addition, we present the cryogenic electron microscopy structure of an Escherichia coli ribosome stalled during translation of speFL in the presence of ornithine. The structure shows how the ribosome and the SpeFL sensor domain form a highly selective binding pocket that accommodates a single ornithine molecule but excludes near-cognate ligands. Ornithine pre-associates with the ribosome and is then held in place by the sensor domain, leading to the compaction of the SpeFL effector domain and blocking the action of release factor 1. Thus, our study not only reveals basic strategies by which nascent peptides assist the ribosome in detecting a specific metabolite, but also provides a framework for assessing how ornithine promotes virulence in several human pathogens. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| History | 
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- Structure visualization
Structure visualization
| Movie | 
 
  Movie viewer | 
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| Structure viewer | Molecule:  Molmil  Jmol/JSmol | 
- Downloads & links
Downloads & links
- Download
Download
| PDBx/mmCIF format |  6tc3.cif.gz | 3.5 MB | Display |  PDBx/mmCIF format | 
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| PDB format |  pdb6tc3.ent.gz | Display |  PDB format | |
| PDBx/mmJSON format |  6tc3.json.gz | Tree view |  PDBx/mmJSON format | |
| Others |  Other downloads | 
-Validation report
| Summary document |  6tc3_validation.pdf.gz | 703.6 KB | Display |  wwPDB validaton report | 
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| Full document |  6tc3_full_validation.pdf.gz | 2.1 MB | Display | |
| Data in XML |  6tc3_validation.xml.gz | 277.7 KB | Display | |
| Data in CIF |  6tc3_validation.cif.gz | 460.7 KB | Display | |
| Arichive directory |  https://data.pdbj.org/pub/pdb/validation_reports/tc/6tc3  ftp://data.pdbj.org/pub/pdb/validation_reports/tc/6tc3 | HTTPS FTP | 
-Related structure data
| Related structure data |  10458MC  6tbvC M: map data used to model this data C: citing same article ( | 
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| Similar structure data | 
- Links
Links
- Assembly
Assembly
| Deposited unit |  
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| 1 | 
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- Components
Components
-RNA chain , 5 types, 5 molecules 16S123S105S1MRN1PTR1    
| #1: RNA chain | Mass: 497404.969 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: GenBank: 1108560344 | 
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| #22: RNA chain | Mass: 939607.250 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) | 
| #23: RNA chain | Mass: 38790.090 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: GenBank: 1726050194 | 
| #54: RNA chain | Mass: 2173.331 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) | 
| #55: RNA chain | Mass: 24582.812 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) | 
-30S ribosomal protein  ... , 20 types, 20 molecules S021S031S041S051S061S071S081S091S101S111S121S131S141S151S161S171S181S191S201S211                   
| #2: Protein | Mass: 26781.670 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P0A7V0 | 
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| #3: Protein | Mass: 26031.316 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P0A7V3 | 
| #4: Protein | Mass: 23514.199 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P0A7V8 | 
| #5: Protein | Mass: 17629.398 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P0A7W1 | 
| #6: Protein | Mass: 15727.512 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P02358 | 
| #7: Protein | Mass: 20055.156 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P02359 | 
| #8: Protein | Mass: 14146.557 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P0A7W7 | 
| #9: Protein | Mass: 14886.270 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P0A7X3 | 
| #10: Protein | Mass: 11755.597 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P0A7R5 | 
| #11: Protein | Mass: 13870.975 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P0A7R9 | 
| #12: Protein | Mass: 13814.249 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P0A7S3 | 
| #13: Protein | Mass: 13128.467 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P0A7S9 | 
| #14: Protein | Mass: 11677.637 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P0AG59 | 
| #15: Protein | Mass: 10290.816 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P0ADZ4 | 
| #16: Protein | Mass: 9207.572 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P0A7T3 | 
| #17: Protein | Mass: 9724.491 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P0AG63 | 
| #18: Protein | Mass: 9005.472 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P0A7T7 | 
| #19: Protein | Mass: 10455.355 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P0A7U3 | 
| #20: Protein | Mass: 9708.464 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P0A7U7 | 
| #21: Protein | Mass: 8524.039 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: P68679 | 
+50S ribosomal protein  ... , 29 types, 29 molecules L021L031L041L051L061L091L311L131L141L151L161L171L181L191L201L211L221L231L241L251L271L281L291L301L321L331L341L351L361                            
-Protein/peptide , 1 types, 1 molecules SPE1
| #53: Protein/peptide | Mass: 4286.883 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: UniProt: A0A4S4NWS2, UniProt: P0DTV7*PLUS | 
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-Non-polymers , 6 types, 2491 molecules 










| #56: Chemical | ChemComp-MG / #57: Chemical | ChemComp-K / #58: Chemical | ChemComp-UNX / #59: Chemical | #60: Chemical | ChemComp-ORN / | #61: Water | ChemComp-HOH / |  | 
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-Details
| Has protein modification | Y | 
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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
Sample preparation
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| Source (natural) | 
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| Buffer solution | pH: 7.5 | ||||||||||||||||||||||||||||||||||||||||||
| Buffer component | 
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| Specimen | Conc.: 0.3 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 R2/2 | ||||||||||||||||||||||||||||||||||||||||||
| Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277.15 K | 
- Electron microscopy imaging
Electron microscopy imaging
| Experimental equipment |  Model: Titan Krios / Image courtesy: FEI Company | 
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| Microscopy | Model: FEI TITAN KRIOS Details: Data were collected at the European Synchrotron Radiation Facility (ESRF CM01 cryo-electron microscope) | 
| Electron gun | Electron source:  FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM | 
| Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 130000 X / Nominal defocus max: -1600 nm / Nominal defocus min: -500 nm / Cs: 2.7 mm | 
| Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Temperature (max): 100 K | 
| Image recording | Average exposure time: 9 sec. / Electron dose: 30 e/Å2 / Detector mode: COUNTING / Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Num. of grids imaged: 1 / Num. of real images: 3620 | 
| Image scans | Movie frames/image: 30 / Used frames/image: 1-30 | 
- Processing
Processing
| EM software | 
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| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||
| Particle selection | Num. of particles selected: 226054 | ||||||||||||||||||||||||||||||||||||
| Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||||||||||||||
| 3D reconstruction | Resolution: 2.7 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 68195 / Algorithm: FOURIER SPACE / Symmetry type: POINT | ||||||||||||||||||||||||||||||||||||
| Atomic model building | Protocol: RIGID BODY FIT / Space: REAL | ||||||||||||||||||||||||||||||||||||
| Atomic model building | PDB-ID: 4YBB Accession code: 4YBB / Source name: PDB / Type: experimental model | 
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