+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-13285 | |||||||||||||||
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Title | free 50S in untreated Mycoplasma pneumoniae cells | |||||||||||||||
Map data | ||||||||||||||||
Sample |
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Keywords | In-cell cryo-electron tomography / sub-tomogram analysis / ribosome / translation intermediate state | |||||||||||||||
Function / homology | Function and homology information large ribosomal subunit / large ribosomal subunit rRNA binding / transferase activity / ribosomal large subunit assembly / cytosolic large ribosomal subunit / tRNA binding / rRNA binding / ribosome / structural constituent of ribosome / translation ...large ribosomal subunit / large ribosomal subunit rRNA binding / transferase activity / ribosomal large subunit assembly / cytosolic large ribosomal subunit / tRNA binding / rRNA binding / ribosome / structural constituent of ribosome / translation / ribonucleoprotein complex / response to antibiotic / cytoplasm Similarity search - Function | |||||||||||||||
Biological species | Mycoplasma pneumoniae M129 (bacteria) / Mycoplasma pneumoniae (strain ATCC 29342 / M129) (bacteria) | |||||||||||||||
Method | subtomogram averaging / cryo EM / Resolution: 9.2 Å | |||||||||||||||
Authors | Xue L / Lenz S | |||||||||||||||
Funding support | Germany, United Kingdom, 4 items
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Citation | Journal: Nature / Year: 2022 Title: 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: 2021 Title: 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 | |||||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_13285.map.gz | 2.9 MB | EMDB map data format | |
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Header (meta data) | emd-13285-v30.xml emd-13285.xml | 50.8 KB 50.8 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_13285_fsc.xml | 7.1 KB | Display | FSC data file |
Images | emd_13285.png | 64 KB | ||
Masks | emd_13285_msk_1.map | 30.5 MB | Mask map | |
Filedesc metadata | emd-13285.cif.gz | 10.6 KB | ||
Others | emd_13285_half_map_1.map.gz emd_13285_half_map_2.map.gz | 23.5 MB 23.4 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-13285 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-13285 | HTTPS FTP |
-Validation report
Summary document | emd_13285_validation.pdf.gz | 870.7 KB | Display | EMDB validaton report |
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Full document | emd_13285_full_validation.pdf.gz | 870.3 KB | Display | |
Data in XML | emd_13285_validation.xml.gz | 13 KB | Display | |
Data in CIF | emd_13285_validation.cif.gz | 17.9 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-13285 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-13285 | HTTPS FTP |
-Related structure data
Related structure data | 7patMC 7oocC 7oodC 7p6zC 7pahC 7paiC 7pajC 7pakC 7palC 7pamC 7panC 7paoC 7paqC 7parC 7pasC 7pauC 7ph9C 7phaC 7phbC 7phcC 7pi8C 7pi9C 7piaC 7pibC 7picC 7pioC 7pipC 7piqC 7pirC 7pisC 7pitC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_13285.map.gz / Format: CCP4 / Size: 30.5 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 2.4 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
File | emd_13285_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_13285_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_13285_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Cryo-electron tomograms of untreated Mycoplasma pneumoniae cells
+Supramolecule #1: Cryo-electron tomograms of untreated Mycoplasma pneumoniae cells
+Macromolecule #1: 50S ribosomal protein L34
+Macromolecule #2: 50S ribosomal protein L35
+Macromolecule #3: 50S ribosomal protein L36
+Macromolecule #4: 50S ribosomal protein L2
+Macromolecule #5: 50S ribosomal protein L3
+Macromolecule #6: 50S ribosomal protein L4
+Macromolecule #7: 50S ribosomal protein L5
+Macromolecule #8: 50S ribosomal protein L6
+Macromolecule #9: 50S ribosomal protein L9
+Macromolecule #10: 50S ribosomal protein L10
+Macromolecule #11: 50S ribosomal protein L11
+Macromolecule #12: 50S ribosomal protein L13
+Macromolecule #13: 50S ribosomal protein L14
+Macromolecule #14: 50S ribosomal protein L15
+Macromolecule #15: 50S ribosomal protein L16
+Macromolecule #16: 50S ribosomal protein L17
+Macromolecule #17: 50S ribosomal protein L18
+Macromolecule #18: 50S ribosomal protein L19
+Macromolecule #19: 50S ribosomal protein L20
+Macromolecule #20: 50S ribosomal protein L21
+Macromolecule #21: 50S ribosomal protein L22
+Macromolecule #22: 50S ribosomal protein L23
+Macromolecule #23: 50S ribosomal protein L24
+Macromolecule #24: 50S ribosomal protein L27
+Macromolecule #25: 50S ribosomal protein L28
+Macromolecule #26: 50S ribosomal protein L29
+Macromolecule #27: 50S ribosomal protein L31
+Macromolecule #28: 50S ribosomal protein L32
+Macromolecule #29: 50S ribosomal protein L33 1
+Macromolecule #30: 23S ribosomal RNA
+Macromolecule #31: 5S ribosomal RNA
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | subtomogram averaging |
Aggregation state | cell |
-Sample preparation
Buffer | pH: 7.4 |
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Vitrification | Cryogen name: ETHANE-PROPANE / Instrument: HOMEMADE PLUNGER Details: back-side blotting for 2-3 seconds before plunging using a manual plunger without an environmental chamber. |
Details | Mycoplasma pneumoniae M129 cells grown on gold Quantifoil grids at 37 Celsius before plunge freezing. |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 3.2 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 3.75 µm / Nominal defocus min: 1.5 µm / Nominal magnification: 81000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |