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Yorodumi- PDB-7ss9: Late translocation intermediate with EF-G partially dissociated (... -
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Basic information
| Entry | Database: PDB / ID: 7ss9 | |||||||||||||||
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| Title | Late translocation intermediate with EF-G partially dissociated (Structure V) | |||||||||||||||
|  Components | 
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|  Keywords | RIBOSOME / EF-G / GTP / Translocation | |||||||||||||||
| Function / homology |  Function and homology information guanosine tetraphosphate binding / stringent response / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / translation elongation factor activity / transcriptional attenuation / endoribonuclease inhibitor activity ...guanosine tetraphosphate binding / stringent response / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / translation elongation factor activity / transcriptional attenuation / endoribonuclease inhibitor activity / positive regulation of ribosome biogenesis / RNA-binding transcription regulator activity / translational termination / 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 / 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 / GTPase activity / mRNA binding / GTP binding / DNA binding / RNA binding / zinc ion binding / membrane / cytoplasm / cytosol Similarity search - Function | |||||||||||||||
| Biological species |   Escherichia coli 113303 (bacteria)   Escherichia coli K-12 (bacteria) | |||||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.9 Å | |||||||||||||||
|  Authors | Carbone, C.E. / Loveland, A.B. / Gamper, H.B. / Hou, Y. / Korostelev, A.A. | |||||||||||||||
| Funding support |  Czech Republic, 4items 
 | |||||||||||||||
|  Citation |  Journal: Nat Commun / Year: 2021 Title: Time-resolved cryo-EM visualizes ribosomal translocation with EF-G and GTP. Authors: Christine E Carbone / Anna B Loveland / Howard B Gamper / Ya-Ming Hou / Gabriel Demo / Andrei A Korostelev /    Abstract: During translation, a conserved GTPase elongation factor-EF-G in bacteria or eEF2 in eukaryotes-translocates tRNA and mRNA through the ribosome. EF-G has been proposed to act as a flexible motor that ...During translation, a conserved GTPase elongation factor-EF-G in bacteria or eEF2 in eukaryotes-translocates tRNA and mRNA through the ribosome. EF-G has been proposed to act as a flexible motor that propels tRNA and mRNA movement, as a rigid pawl that biases unidirectional translocation resulting from ribosome rearrangements, or by various combinations of motor- and pawl-like mechanisms. Using time-resolved cryo-EM, we visualized GTP-catalyzed translocation without inhibitors, capturing elusive structures of ribosome•EF-G intermediates at near-atomic resolution. Prior to translocation, EF-G binds near peptidyl-tRNA, while the rotated 30S subunit stabilizes the EF-G GTPase center. Reverse 30S rotation releases Pi and translocates peptidyl-tRNA and EF-G by ~20 Å. An additional 4-Å translocation initiates EF-G dissociation from a transient ribosome state with highly swiveled 30S head. The structures visualize how nearly rigid EF-G rectifies inherent and spontaneous ribosomal dynamics into tRNA-mRNA translocation, whereas GTP hydrolysis and Pi release drive EF-G dissociation. | |||||||||||||||
| History | 
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- Structure visualization
Structure visualization
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| Structure viewer | Molecule:  Molmil  Jmol/JSmol | 
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| PDBx/mmCIF format |  7ss9.cif.gz | 3.2 MB | Display |  PDBx/mmCIF format | 
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| PDB format |  pdb7ss9.ent.gz | Display |  PDB format | |
| PDBx/mmJSON format |  7ss9.json.gz | Tree view |  PDBx/mmJSON format | |
| Others |  Other downloads | 
-Validation report
| Summary document |  7ss9_validation.pdf.gz | 1.7 MB | Display |  wwPDB validaton report | 
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| Full document |  7ss9_full_validation.pdf.gz | 1.8 MB | Display | |
| Data in XML |  7ss9_validation.xml.gz | 214.9 KB | Display | |
| Data in CIF |  7ss9_validation.cif.gz | 374.1 KB | Display | |
| Arichive directory |  https://data.pdbj.org/pub/pdb/validation_reports/ss/7ss9  ftp://data.pdbj.org/pub/pdb/validation_reports/ss/7ss9 | HTTPS FTP | 
-Related structure data
| Related structure data |  25405MC  7ssdC  7sslC  7ssnC  7ssoC  7sswC  7st2C  7st6C  7st7C M: map data used to model this data C: citing same article ( | 
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| Similar structure data | 
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- Assembly
Assembly
| Deposited unit |  
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| 1 | 
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- Components
Components
-RNA chain , 6 types, 6 molecules 312645     
| #1: RNA chain | Mass: 498725.406 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: GenBank: 1126835768 | 
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| #2: RNA chain | Mass: 941305.250 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: GenBank: 802133627 | 
| #3: RNA chain | Mass: 38813.133 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: GenBank: 1266961702 | 
| #5: RNA chain | Mass: 24802.785 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / References: GenBank: 1275521620 | 
| #36: RNA chain | Mass: 12710.771 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K-12 | 
| #37: RNA chain | Mass: 24862.746 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: MRE600 / References: GenBank: 1848949880 | 
-Protein , 1 types, 1 molecules 8
| #4: Protein | Mass: 76902.242 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.)   Escherichia coli 113303 (bacteria) / Gene: fusA, HMPREF1591_03890 Production host:   Escherichia coli 'BL21-Gold(DE3)pLysS AG' (bacteria) References: UniProt: U9XYS4 | 
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+50S ribosomal protein  ... , 30 types, 30 molecules bcdefgaijklmnopqrstuvwxyzBCDEF                             
-30S ribosomal protein  ... , 20 types, 20 molecules GHIJKLMNOPQRSTUVWXYZ                   
| #38: Protein | Mass: 25015.816 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P0A7V0 | 
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| #39: Protein | Mass: 23078.785 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P0A7V3 | 
| #40: Protein | Mass: 23383.002 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P0A7V8 | 
| #41: Protein | Mass: 16532.088 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P0A7W1 | 
| #42: Protein | Mass: 11669.371 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P02358 | 
| #43: Protein | Mass: 16861.523 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P02359 | 
| #44: Protein | Mass: 14015.361 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P0A7W7 | 
| #45: Protein | Mass: 14554.882 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P0A7X3 | 
| #46: Protein | Mass: 11196.988 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P0A7R5 | 
| #47: Protein | Mass: 12388.068 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P0A7R9 | 
| #48: Protein | Mass: 13636.961 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P0A7S3 | 
| #49: Protein | Mass: 12625.753 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P0A7S9 | 
| #50: Protein | Mass: 11475.364 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P0AG59 | 
| #51: Protein | Mass: 10159.621 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P0ADZ4 | 
| #52: Protein | Mass: 9207.572 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P0A7T3 | 
| #53: Protein | Mass: 9263.946 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P0AG63 | 
| #54: Protein | Mass: 7606.768 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P0A7T7 | 
| #55: Protein | Mass: 9057.626 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P0A7U3 | 
| #56: Protein | Mass: 9506.190 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P0A7U7 | 
| #57: Protein | Mass: 7763.073 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli K-12 (bacteria) / Strain: K12 / References: UniProt: P68679 | 
-Details
| Has ligand of interest | Y | 
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| Has protein modification | Y | 
-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
| Component | Name: Late translocation intermediate with EF-G partially dissociated (Structure V) Type: RIBOSOME / Entity ID: all / Source: MULTIPLE SOURCES | 
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| Molecular weight | Value: 2.7 MDa / Experimental value: NO | 
| Source (natural) | Organism:   Escherichia coli K-12 (bacteria) | 
| Source (recombinant) | Organism:   Escherichia coli 'BL21-Gold(DE3)pLysS AG' (bacteria) | 
| 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
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: 800 nm | 
| Image recording | Electron dose: 47.6 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) | 
- Processing
Processing
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| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||||||||||||||
| 3D reconstruction | Resolution: 3.9 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 1105 / Symmetry type: POINT | ||||||||||||||||||||||||||||||||||||||||||||||||
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