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Yorodumi- PDB-7pam: 70S ribosome with A*- and P/E-site tRNAs in Mycoplasma pneumoniae... -
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Open data
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Basic information
| Entry | Database: PDB / ID: 7pam | |||||||||||||||
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| Title | 70S ribosome with A*- and P/E-site tRNAs in Mycoplasma pneumoniae cells | |||||||||||||||
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Keywords | TRANSLATION / In-cell cryo-electron tomography / sub-tomogram analysis / ribosome / translation intermediate state | |||||||||||||||
| Function / homology | Function and homology informationlarge ribosomal subunit / 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 ...large ribosomal subunit / 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 / ribonucleoprotein complex / response to antibiotic / mRNA binding / RNA binding / zinc ion binding / cytoplasm / cytosol Similarity search - Function | |||||||||||||||
| Biological species | Mycoplasma pneumoniae M129 (bacteria) Mycoplasma pneumoniae (Filterable agent of primary atypical pneumonia) | |||||||||||||||
| Method | ELECTRON MICROSCOPY / subtomogram averaging / cryo EM / Resolution: 6.8 Å | |||||||||||||||
Authors | Xue, L. / Lenz, S. / Rappsilber, J. / Mahamid, J. | |||||||||||||||
| Funding support | Germany, United Kingdom, 4items
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Citation | Journal: Nature / Year: 2022Title: Visualizing translation dynamics at atomic detail inside a bacterial cell. Authors: Liang Xue / Swantje Lenz / Maria Zimmermann-Kogadeeva / Dimitry Tegunov / Patrick Cramer / Peer Bork / Juri Rappsilber / Julia Mahamid / ![]() Abstract: Translation is the fundamental process of protein synthesis and is catalysed by the ribosome in all living cells. Here we use advances in cryo-electron tomography and sub-tomogram analysis to ...Translation is the fundamental process of protein synthesis and is catalysed by the ribosome in all living cells. Here we use advances in cryo-electron tomography and sub-tomogram analysis to visualize the structural dynamics of translation inside the bacterium Mycoplasma pneumoniae. To interpret the functional states in detail, we first obtain a high-resolution in-cell average map of all translating ribosomes and build an atomic model for the M. pneumoniae ribosome that reveals distinct extensions of ribosomal proteins. Classification then resolves 13 ribosome states that differ in their conformation and composition. These recapitulate major states that were previously resolved in vitro, and reflect intermediates during active translation. On the basis of these states, we animate translation elongation inside native cells and show how antibiotics reshape the cellular translation landscapes. During translation elongation, ribosomes often assemble in defined three-dimensional arrangements to form polysomes. By mapping the intracellular organization of translating ribosomes, we show that their association into polysomes involves a local coordination mechanism that is mediated by the ribosomal protein L9. We propose that an extended conformation of L9 within polysomes mitigates collisions to facilitate translation fidelity. Our work thus demonstrates the feasibility of visualizing molecular processes at atomic detail inside cells. #1: Journal: Biorxiv / Year: 2021Title: Visualizing translation dynamics at atomic detail inside a bacterial cell Authors: Xue, L. / Lenz, S. / Zimmermann-Kogadeeva, M. / Tegunov, D. / Cramer, P. / Bork, P. / Rappsilber, J. / Mahamid, J. | |||||||||||||||
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Structure visualization
| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 7pam.cif.gz | 3.2 MB | Display | PDBx/mmCIF format |
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| PDB format | pdb7pam.ent.gz | Display | PDB format | |
| PDBx/mmJSON format | 7pam.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Summary document | 7pam_validation.pdf.gz | 1.8 MB | Display | wwPDB validaton report |
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| Full document | 7pam_full_validation.pdf.gz | 1.9 MB | Display | |
| Data in XML | 7pam_validation.xml.gz | 205.4 KB | Display | |
| Data in CIF | 7pam_validation.cif.gz | 356.3 KB | Display | |
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/pa/7pam ftp://data.pdbj.org/pub/pdb/validation_reports/pa/7pam | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 13277MC ![]() 7oocC ![]() 7oodC ![]() 7p6zC ![]() 7pahC ![]() 7paiC ![]() 7pajC ![]() 7pakC ![]() 7palC ![]() 7panC ![]() 7paoC ![]() 7paqC ![]() 7parC ![]() 7pasC ![]() 7patC ![]() 7pauC ![]() 7ph9C ![]() 7phaC ![]() 7phbC ![]() 7phcC ![]() 7pi8C ![]() 7pi9C ![]() 7piaC ![]() 7pibC ![]() 7picC ![]() 7pioC ![]() 7pipC ![]() 7piqC ![]() 7pirC ![]() 7pisC ![]() 7pitC M: map data used to model this data C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
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Assembly
| Deposited unit | ![]()
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Components
+50S ribosomal protein ... , 29 types, 29 molecules 012abcdefghijklmnopqrstuvwxyz
-30S ribosomal protein ... , 20 types, 20 molecules ABCDEFGHIJKLMNOPQRST
| #4: Protein | Mass: 33468.629 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: P75560 |
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| #5: Protein | Mass: 30657.205 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: P41205 |
| #6: Protein | Mass: 23817.645 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: P46775 |
| #7: Protein | Mass: 24138.008 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: Q50301 |
| #8: Protein | Mass: 25430.465 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: P75543 |
| #9: Protein | Mass: 17897.029 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: P75545 |
| #10: Protein | Mass: 15903.083 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: Q50304 |
| #11: Protein | Mass: 15149.735 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: P75179 |
| #12: Protein | Mass: 12226.571 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: P75581 |
| #13: Protein | Mass: 12709.851 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: Q50296 |
| #14: Protein | Mass: 15666.580 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: P75546 |
| #15: Protein | Mass: 14209.607 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: Q50297 |
| #16: Protein | Mass: 6901.364 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: Q50305 |
| #17: Protein | Mass: 9921.687 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: P75173 |
| #18: Protein | Mass: 10806.921 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / References: UniProt: A0A0H3DLS7 |
| #19: Protein | Mass: 9859.643 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: Q50309 |
| #20: Protein | Mass: 12411.522 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: P75541 |
| #21: Protein | Mass: 10057.626 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: P75576 |
| #22: Protein | Mass: 9993.599 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: P75237 |
| #23: Protein | Mass: 7539.182 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) / Strain: ATCC 29342 / M129 / References: UniProt: P57079 |
-RNA chain , 4 types, 5 molecules 34568
| #50: RNA chain | Mass: 940911.500 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) |
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| #51: RNA chain | Mass: 34796.676 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) |
| #52: RNA chain | Mass: 491243.625 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) |
| #53: RNA chain | Mass: 24470.521 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Mycoplasma pneumoniae M129 (bacteria) |
-Details
| Has protein modification | Y |
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-Experimental details
-Experiment
| Experiment | Method: ELECTRON MICROSCOPY |
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| EM experiment | Aggregation state: CELL / 3D reconstruction method: subtomogram averaging |
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Sample preparation
| Component | Name: Cryo-electron tomograms of untreated Mycoplasma pneumoniae cells Type: CELL Details: Ribosome sub-tomograms extracted in silico from tomograms of intact cell, refinement with a 70S mask Entity ID: all / Source: NATURAL |
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| Source (natural) | Organism: Mycoplasma pneumoniae M129 (bacteria) |
| Buffer solution | pH: 7.4 |
| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES Details: Mycoplasma pneumoniae M129 cells grown on gold Quantifoil grids at 37 Celsius before plunge freezing. |
| Vitrification | Instrument: HOMEMADE PLUNGER / Cryogen name: ETHANE-PROPANE Details: back-side blotting for 2-3 seconds before plunging using a manual plunger without an environmental chamber |
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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 / Nominal magnification: 81000 X / Nominal defocus max: 3750 nm / Nominal defocus min: 1500 nm / Cs: 2.7 mm |
| Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
| Image recording | Electron dose: 3.2 e/Å2 / Detector mode: COUNTING / Film or detector model: GATAN K2 SUMMIT (4k x 4k) |
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Processing
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| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | |||||||||||||||||||||||||||||||||||||||||||||
| Symmetry | Point symmetry: C1 (asymmetric) | |||||||||||||||||||||||||||||||||||||||||||||
| 3D reconstruction | Resolution: 6.8 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 6587 / Symmetry type: POINT | |||||||||||||||||||||||||||||||||||||||||||||
| EM volume selection | Num. of tomograms: 356 / Num. of volumes extracted: 77539 | |||||||||||||||||||||||||||||||||||||||||||||
| Atomic model building | Protocol: FLEXIBLE FIT / Space: REAL | |||||||||||||||||||||||||||||||||||||||||||||
| Atomic model building | 3D fitting-ID: 1 / Source name: PDB / Type: experimental model
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About Yorodumi



Mycoplasma pneumoniae M129 (bacteria)
Germany,
United Kingdom, 4items
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FIELD EMISSION GUN
