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Yorodumi- PDB-7n2v: Elongating 70S ribosome complex in a spectinomycin-stalled interm... -
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Basic information
| Entry | Database: PDB / ID: 7n2v | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Title | Elongating 70S ribosome complex in a spectinomycin-stalled intermediate state of translocation bound to EF-G in an active, GTP conformation (INT1) | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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 Keywords | RIBOSOME / Elongation complex / tRNA / mRNA / translocation / EF-G & GTP | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Function / homology |  Function and homology informationribosome disassembly / guanosine tetraphosphate binding / negative regulation of cytoplasmic translational initiation / stringent response / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / translation elongation factor activity / transcriptional attenuation / endoribonuclease inhibitor activity / positive regulation of ribosome biogenesis ...ribosome disassembly / guanosine tetraphosphate binding / negative regulation of cytoplasmic translational initiation / stringent response / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / 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 / 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 / GTPase activity / mRNA binding / GTP binding / DNA binding / RNA binding / zinc ion binding / membrane / cytoplasm / cytosol Similarity search - Function  | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Biological species | ![]() ![]()  | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.54 Å | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 Authors | Rundlet, E.J. / Holm, M. / Schacherl, M. / Natchiar, K.S. / Altman, R.B. / Spahn, C.M.T. / Myasnikov, A.G. / Blanchard, S.C. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 Citation |  Journal: Nature / Year: 2021Title: Structural basis of early translocation events on the ribosome. Authors: Emily J Rundlet / Mikael Holm / Magdalena Schacherl / S Kundhavai Natchiar / Roger B Altman / Christian M T Spahn / Alexander G Myasnikov / Scott C Blanchard /   ![]() Abstract: Peptide-chain elongation during protein synthesis entails sequential aminoacyl-tRNA selection and translocation reactions that proceed rapidly (2-20 per second) and with a low error rate (around ...Peptide-chain elongation during protein synthesis entails sequential aminoacyl-tRNA selection and translocation reactions that proceed rapidly (2-20 per second) and with a low error rate (around 10 to 10 at each step) over thousands of cycles. The cadence and fidelity of ribosome transit through mRNA templates in discrete codon increments is a paradigm for movement in biological systems that must hold for diverse mRNA and tRNA substrates across domains of life. Here we use single-molecule fluorescence methods to guide the capture of structures of early translocation events on the bacterial ribosome. Our findings reveal that the bacterial GTPase elongation factor G specifically engages spontaneously achieved ribosome conformations while in an active, GTP-bound conformation to unlock and initiate peptidyl-tRNA translocation. These findings suggest that processes intrinsic to the pre-translocation ribosome complex can regulate the rate of protein synthesis, and that energy expenditure is used later in the translocation mechanism than previously proposed.  | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Structure visualization
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| Structure viewer | Molecule:  Molmil Jmol/JSmol | 
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Downloads & links
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Download
| PDBx/mmCIF format |  7n2v.cif.gz | 3.4 MB | Display |  PDBx/mmCIF format | 
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| PDB format |  pdb7n2v.ent.gz | Display |  PDB format | |
| PDBx/mmJSON format |  7n2v.json.gz | Tree view |  PDBx/mmJSON format | |
| Others |  Other downloads | 
-Validation report
| Summary document |  7n2v_validation.pdf.gz | 2.2 MB | Display |  wwPDB validaton report | 
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| Full document |  7n2v_full_validation.pdf.gz | 2.3 MB | Display | |
| Data in XML |  7n2v_validation.xml.gz | 246.5 KB | Display | |
| Data in CIF |  7n2v_validation.cif.gz | 423.9 KB | Display | |
| Arichive directory |  https://data.pdbj.org/pub/pdb/validation_reports/n2/7n2v ftp://data.pdbj.org/pub/pdb/validation_reports/n2/7n2v | HTTPS FTP  | 
-Related structure data
| Related structure data | ![]() 24134MC ![]() 7n1pC ![]() 7n2cC ![]() 7n2uC ![]() 7n30C ![]() 7n31C M: map data used to model this data C: citing same article (  | 
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| Similar structure data | 
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Links
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Assembly
| Deposited unit | ![]() 
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| 1 | 
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Components
-RNA chain , 6 types, 6 molecules 16mR235PtDt     
| #1: RNA chain |   Mass: 497391.906 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: 16S rRNA / Source: (natural)  ![]()  | 
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| #22: RNA chain |   Mass: 19128.443 Da / Num. of mol.: 1 / Source method: obtained synthetically / Details: mRNA / Source: (synth.)  ![]()  | 
| #23: RNA chain |   Mass: 941812.562 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: 23S rRNA / Source: (natural)  ![]()  | 
| #24: RNA chain |   Mass: 38790.090 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: 5S rRNA / Source: (natural)  ![]()  | 
| #58: RNA chain |   Mass: 34369.590 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: tRNA / Source: (gene. exp.) ![]() ![]()  | 
| #59: RNA chain |   Mass: 34451.734 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: tRNA / Source: (gene. exp.) ![]() ![]()  | 
-30S ribosomal protein  ... , 20 types, 20 molecules SBSCSDSESFSGSHSISJSKSLSMSNSOSPSQSRSSSTSU                   
| #2: Protein |   Mass: 26781.670 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S2 / Source: (natural)  ![]()  | 
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| #3: Protein |   Mass: 26031.316 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S3 / Source: (natural)  ![]()  | 
| #4: Protein |   Mass: 23514.199 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S4 / Source: (natural)  ![]()  | 
| #5: Protein |   Mass: 17629.398 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S5 / Source: (natural)  ![]()  | 
| #6: Protein |   Mass: 15727.512 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S6 / Source: (natural)  ![]()  | 
| #7: Protein |   Mass: 20055.156 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S7 / Source: (natural)  ![]()  | 
| #8: Protein |   Mass: 14146.557 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S8 / Source: (natural)  ![]()  | 
| #9: Protein |   Mass: 14886.270 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S9 / Source: (natural)  ![]()  | 
| #10: Protein |   Mass: 11755.597 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S10 / Source: (natural)  ![]()  | 
| #11: Protein |   Mass: 13870.975 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S11 / Source: (natural)  ![]()  | 
| #12: Protein |   Mass: 13814.249 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S12 / Source: (natural)  ![]()  | 
| #13: Protein |   Mass: 13128.467 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S13 / Source: (natural)  ![]()  | 
| #14: Protein |   Mass: 11606.560 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S14 / Source: (natural)  ![]()  | 
| #15: Protein |   Mass: 10290.816 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S15 / Source: (natural)  ![]()  | 
| #16: Protein |   Mass: 9207.572 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S16 / Source: (natural)  ![]()  | 
| #17: Protein |   Mass: 9724.491 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S17 / Source: (natural)  ![]()  | 
| #18: Protein |   Mass: 9005.472 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S18 / Source: (natural)  ![]()  | 
| #19: Protein |   Mass: 10455.355 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S19 / Source: (natural)  ![]()  | 
| #20: Protein |   Mass: 9708.464 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S20 / Source: (natural)  ![]()  | 
| #21: Protein |   Mass: 8524.039 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Ribosomal Protein S21 / Source: (natural)  ![]()  | 
+50S ribosomal protein  ... , 31 types, 31 molecules LBLCLDLELFLILJLKLMLNLOLPLQLRLSLTLULVLWLXLYLaLbLcLdLeLfLgLhLiLj                              
-Protein / Protein/peptide , 2 types, 2 molecules EFPp 
| #56: Protein |   Mass: 77632.109 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: Elongation factor (EF-G) / Source: (gene. exp.) ![]() ![]()  | 
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| #57: Protein/peptide |   Mass: 425.565 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Nascent peptide / Source: (natural)  ![]()  | 
-Non-polymers , 8 types, 398 molecules 














| #60: Chemical | | #61: Chemical | ChemComp-PUT / #62: Chemical | ChemComp-MG / #63: Chemical | #64: Chemical | #65: Chemical | #66: Chemical |  ChemComp-GTP /  | #67: Water |  ChemComp-HOH /  |  | 
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-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 | 
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Sample preparation
| Component | Name: Elongating 70S ribosome complex in a spectinomycin-stalled intermediate state of translocation bound to EF-G in an active, GTP conformation (INT1) Type: RIBOSOME / Entity ID: #1-#56, #58, #57, #59 / Source: NATURAL  | 
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| Source (natural) | Organism: ![]()  | 
| Buffer solution | pH: 7.5 | 
| Specimen | Conc.: 5.5 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 | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 95 % / Chamber temperature: 283 K | 
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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 | 
| Image recording | Electron dose: 87 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) | 
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Processing
| CTF correction | Type: PHASE FLIPPING ONLY | 
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| 3D reconstruction | Resolution: 2.54 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 33688 / Symmetry type: POINT | 
| Atomic model building | Protocol: OTHER / Space: RECIPROCAL | 
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