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Yorodumi- EMDB-8620: 70S ribosome bound with near-cognate ternary complex base-paired ... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-8620 | |||||||||||||||
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Title | 70S ribosome bound with near-cognate ternary complex base-paired to A site codon, closed 30S (Structure III-nc) | |||||||||||||||
Map data | 70S ribosome bound with near-cognate ternary complex base-paired to A site codon, closed 30S (Structure III-nc): filtered map from Frealign (FFILT setting in Frealign = T), used for model building after B-factor sharpening. This volume not B-factor sharpened. | |||||||||||||||
Sample |
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Keywords | Ribosome / EF-Tu / tRNA | |||||||||||||||
Function / homology | Function and homology information guanyl-nucleotide exchange factor complex / negative regulation of cytoplasmic translational initiation / guanosine tetraphosphate binding / stringent response / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / translational elongation ...guanyl-nucleotide exchange factor complex / negative regulation of cytoplasmic translational initiation / guanosine tetraphosphate binding / stringent response / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / translational elongation / transcriptional attenuation / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity / positive regulation of ribosome biogenesis / negative regulation of cytoplasmic translation / translation elongation factor activity / four-way junction DNA binding / translational termination / DnaA-L2 complex / translation repressor activity / negative regulation of DNA-templated DNA replication initiation / negative regulation of translational initiation / regulation of mRNA stability / mRNA regulatory element binding translation repressor activity / ribosome assembly / assembly of large subunit precursor of preribosome / positive regulation of RNA splicing / transcription elongation factor complex / cytosolic ribosome assembly / regulation of DNA-templated transcription elongation / DNA endonuclease activity / response to reactive oxygen species / transcription antitermination / regulation of cell growth / translational initiation / DNA-templated transcription termination / maintenance of translational fidelity / response to radiation / mRNA 5'-UTR binding / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / large ribosomal subunit / ribosome biogenesis / ribosome binding / regulation of translation / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / large ribosomal subunit rRNA binding / transferase activity / cytosolic small ribosomal subunit / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / tRNA binding / molecular adaptor activity / negative regulation of translation / rRNA binding / ribosome / structural constituent of 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 / plasma membrane / cytosol / cytoplasm Similarity search - Function | |||||||||||||||
Biological species | Escherichia coli (strain K12) (bacteria) | |||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.8 Å | |||||||||||||||
Authors | Loveland AB / Demo G | |||||||||||||||
Funding support | United States, 4 items
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Citation | Journal: Nature / Year: 2017 Title: Ensemble cryo-EM elucidates the mechanism of translation fidelity. Authors: Anna B Loveland / Gabriel Demo / Nikolaus Grigorieff / Andrei A Korostelev / Abstract: Gene translation depends on accurate decoding of mRNA, the structural mechanism of which remains poorly understood. Ribosomes decode mRNA codons by selecting cognate aminoacyl-tRNAs delivered by ...Gene translation depends on accurate decoding of mRNA, the structural mechanism of which remains poorly understood. Ribosomes decode mRNA codons by selecting cognate aminoacyl-tRNAs delivered by elongation factor Tu (EF-Tu). Here we present high-resolution structural ensembles of ribosomes with cognate or near-cognate aminoacyl-tRNAs delivered by EF-Tu. Both cognate and near-cognate tRNA anticodons explore the aminoacyl-tRNA-binding site (A site) of an open 30S subunit, while inactive EF-Tu is separated from the 50S subunit. A transient conformation of decoding-centre nucleotide G530 stabilizes the cognate codon-anticodon helix, initiating step-wise 'latching' of the decoding centre. The resulting closure of the 30S subunit docks EF-Tu at the sarcin-ricin loop of the 50S subunit, activating EF-Tu for GTP hydrolysis and enabling accommodation of the aminoacyl-tRNA. By contrast, near-cognate complexes fail to induce the G530 latch, thus favouring open 30S pre-accommodation intermediates with inactive EF-Tu. This work reveals long-sought structural differences between the pre-accommodation of cognate and near-cognate tRNAs that elucidate the mechanism of accurate decoding. | |||||||||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_8620.map.gz | 100.7 MB | EMDB map data format | |
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Header (meta data) | emd-8620-v30.xml emd-8620.xml | 85.5 KB 85.5 KB | Display Display | EMDB header |
Images | emd_8620.png | 164.3 KB | ||
Filedesc metadata | emd-8620.cif.gz | 15.3 KB | ||
Others | emd_8620_half_map_1.map.gz emd_8620_half_map_2.map.gz | 101.6 MB 101.6 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-8620 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-8620 | HTTPS FTP |
-Validation report
Summary document | emd_8620_validation.pdf.gz | 902.3 KB | Display | EMDB validaton report |
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Full document | emd_8620_full_validation.pdf.gz | 901.8 KB | Display | |
Data in XML | emd_8620_validation.xml.gz | 18.5 KB | Display | |
Data in CIF | emd_8620_validation.cif.gz | 22 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-8620 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-8620 | HTTPS FTP |
-Related structure data
Related structure data | 5uyqMC 8615C 8616C 8617C 8618C 8619C 5uykC 5uylC 5uymC 5uynC 5uypC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_8620.map.gz / Format: CCP4 / Size: 421.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | 70S ribosome bound with near-cognate ternary complex base-paired to A site codon, closed 30S (Structure III-nc): filtered map from Frealign (FFILT setting in Frealign = T), used for model building after B-factor sharpening. This volume not B-factor sharpened. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.82 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Half map: 70S ribosome bound with near-cognate ternary complex base-paired...
File | emd_8620_half_map_1.map | ||||||||||||
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Annotation | 70S ribosome bound with near-cognate ternary complex base-paired to A site codon, closed 30S (Structure III-nc): half map (1) suitable for resolution calculation. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: 70S ribosome bound with near-cognate ternary complex base-paired...
File | emd_8620_half_map_2.map | ||||||||||||
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Annotation | 70S ribosome bound with near-cognate ternary complex base-paired to A site codon, closed 30S (Structure III-nc): half map (2) suitable for resolution calculation. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : 70S ribosome bound with near-cognate ternary complex base-paired ...
+Supramolecule #1: 70S ribosome bound with near-cognate ternary complex base-paired ...
+Macromolecule #1: 50S ribosomal protein L2
+Macromolecule #2: 50S ribosomal protein L3
+Macromolecule #3: 50S ribosomal protein L4
+Macromolecule #4: 50S ribosomal protein L5
+Macromolecule #5: 50S ribosomal protein L6
+Macromolecule #6: 50S ribosomal protein L9
+Macromolecule #7: 50S ribosomal protein L10
+Macromolecule #8: 50S ribosomal protein L11
+Macromolecule #9: 50S ribosomal protein L13
+Macromolecule #10: 50S ribosomal protein L14
+Macromolecule #11: 50S ribosomal protein L15
+Macromolecule #12: 50S ribosomal protein L16
+Macromolecule #13: 50S ribosomal protein L17
+Macromolecule #14: 50S ribosomal protein L18
+Macromolecule #15: 50S ribosomal protein L19
+Macromolecule #16: 50S ribosomal protein L20
+Macromolecule #17: 50S ribosomal protein L21
+Macromolecule #18: 50S ribosomal protein L22
+Macromolecule #19: 50S ribosomal protein L23
+Macromolecule #20: 50S ribosomal protein L24
+Macromolecule #21: 50S ribosomal protein L25
+Macromolecule #22: 50S ribosomal protein L27
+Macromolecule #23: 50S ribosomal protein L28
+Macromolecule #24: 50S ribosomal protein L29
+Macromolecule #25: 50S ribosomal protein L30
+Macromolecule #26: 50S ribosomal protein L31
+Macromolecule #27: 50S ribosomal protein L32
+Macromolecule #28: 50S ribosomal protein L33
+Macromolecule #29: 50S ribosomal protein L34
+Macromolecule #30: 50S ribosomal protein L35
+Macromolecule #31: 50S ribosomal protein L36
+Macromolecule #32: 30S ribosomal protein S2
+Macromolecule #33: 30S ribosomal protein S3
+Macromolecule #34: 30S ribosomal protein S4
+Macromolecule #35: 30S ribosomal protein S5
+Macromolecule #36: 30S ribosomal protein S6
+Macromolecule #37: 30S ribosomal protein S7
+Macromolecule #38: 30S ribosomal protein S8
+Macromolecule #39: 30S ribosomal protein S9
+Macromolecule #40: 30S ribosomal protein S10
+Macromolecule #41: 30S ribosomal protein S11
+Macromolecule #42: 30S ribosomal protein S12
+Macromolecule #43: 30S ribosomal protein S13
+Macromolecule #44: 30S ribosomal protein S14
+Macromolecule #45: 30S ribosomal protein S15
+Macromolecule #46: 30S ribosomal protein S16
+Macromolecule #47: 30S ribosomal protein S17
+Macromolecule #48: 30S ribosomal protein S18
+Macromolecule #49: 30S ribosomal protein S19
+Macromolecule #50: 30S ribosomal protein S20
+Macromolecule #51: 30S ribosomal protein S21
+Macromolecule #52: 50S ribosomal protein L1
+Macromolecule #59: Elongation factor Tu 2
+Macromolecule #53: 16S ribosomal RNA
+Macromolecule #54: 23S ribosomal RNA
+Macromolecule #55: 5S ribosomal RNA
+Macromolecule #56: tRNAfMet
+Macromolecule #57: mRNA
+Macromolecule #58: tRNALys
+Macromolecule #60: N-FORMYLMETHIONINE
+Macromolecule #61: LYSINE
+Macromolecule #62: MAGNESIUM ION
+Macromolecule #63: PHOSPHOMETHYLPHOSPHONIC ACID GUANYLATE ESTER
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 Component:
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Grid | Model: C-flat-1.2/1.3 / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: HOLEY ARRAY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 45 sec. / Pretreatment - Atmosphere: AIR | |||||||||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 275 K / Instrument: GATAN CRYOPLUNGE 3 Details: 2 uL of complex was applied to each grid. After a 10-second incubation, the grids were blotted for 2 to 4 seconds.. | |||||||||||||||||||||
Details | 125 nM 50S, 125 nM 30S, 625 nM mRNA, 250 nM fMet-tRNAfMet, 1.25 micromolar EF-Tu, 500 micromolar GDPCP, 1.25 micromolar Lys-tRNALys |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Digitization - Dimensions - Width: 7676 pixel / Digitization - Dimensions - Height: 7420 pixel / Digitization - Frames/image: 1-30 / Number grids imaged: 3 / Number real images: 1773 / Average exposure time: 0.4 sec. / Average electron dose: 1.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Calibrated magnification: 60976 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 5.0 µm / Nominal defocus min: 0.5 µm / Nominal magnification: 60976 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
+Image processing
-Atomic model buiding 1
Refinement | Space: REAL / Protocol: FLEXIBLE FIT / Target criteria: correlation coefficient |
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Output model | PDB-5uyq: |