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Yorodumi- EMDB-29820: Structure of the Escherichia coli 70S ribosome in complex with EF... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-29820 | |||||||||
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Title | Structure of the Escherichia coli 70S ribosome in complex with EF-Tu and Ile-tRNAIle(LAU) bound to the near-cognate AUG codon (Structure II) | |||||||||
Map data | Ribosome bound to EF-Tu-GDPCP in the presence of paromomycin with AUG codon in the A site, A-site Ile-tRNAIle(LAU), P-site tRNAfMet and E-site tRNAIle(LAU)(Structure II) | |||||||||
Sample |
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Keywords | Elongation factor Tu / paromomycin / lysidine 34 / cryo-EM / tRNA / RIBOSOME | |||||||||
Function / homology | Function and homology information negative regulation of cytoplasmic translational initiation / guanosine tetraphosphate binding / stringent response / positive regulation of ribosome biogenesis / translation elongation factor activity / translational termination / DnaA-L2 complex / negative regulation of translational initiation / negative regulation of DNA-templated DNA replication initiation / mRNA regulatory element binding translation repressor activity ...negative regulation of cytoplasmic translational initiation / guanosine tetraphosphate binding / stringent response / positive regulation of ribosome biogenesis / translation elongation factor activity / translational termination / DnaA-L2 complex / negative regulation of translational initiation / negative regulation of DNA-templated DNA replication initiation / mRNA regulatory element binding translation repressor activity / ribosome assembly / assembly of large subunit precursor of preribosome / cytosolic ribosome assembly / transcription antitermination / regulation of cell growth / translational initiation / DNA-templated transcription termination / maintenance of translational fidelity / mRNA 5'-UTR binding / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / large ribosomal subunit / ribosome binding / ribosomal small subunit assembly / large ribosomal subunit rRNA binding / 5S rRNA binding / transferase activity / small ribosomal subunit / cytosolic small ribosomal subunit / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / tRNA binding / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / translation / ribonucleoprotein complex / response to antibiotic / GTPase activity / mRNA binding / GTP binding / RNA binding / zinc ion binding / membrane / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Escherichia coli (E. coli) / Escherichia phage T4 (virus) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.1 Å | |||||||||
Authors | Rybak MY / Gagnon MG | |||||||||
Funding support | United States, 2 items
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Citation | Journal: Nat Struct Mol Biol / Year: 2024 Title: Structures of the ribosome bound to EF-Tu-isoleucine tRNA elucidate the mechanism of AUG avoidance. Authors: Mariia Yu Rybak / Matthieu G Gagnon / Abstract: The frequency of errors upon decoding of messenger RNA by the bacterial ribosome is low, with one misreading event per 1 × 10 codons. In the universal genetic code, the AUN codon box specifies ...The frequency of errors upon decoding of messenger RNA by the bacterial ribosome is low, with one misreading event per 1 × 10 codons. In the universal genetic code, the AUN codon box specifies two amino acids, isoleucine and methionine. In bacteria and archaea, decoding specificity of the AUA and AUG codons relies on the wobble avoidance strategy that requires modification of C34 in the anticodon loop of isoleucine transfer RNA (tRNA). Bacterial tRNA with 2-lysylcytidine (lysidine) at the wobble position deciphers AUA while avoiding AUG. Here we report cryo-electron microscopy structures of the Escherichia coli 70S ribosome complexed with elongation factor thermo unstable (EF-Tu) and isoleucine-tRNA in the process of decoding AUA and AUG. Lysidine in tRNA excludes AUG by promoting the formation of an unusual Hoogsteen purine-pyrimidine nucleobase geometry at the third position of the codon, weakening the interactions with the mRNA and destabilizing the EF-Tu ternary complex. Our findings elucidate the molecular mechanism by which tRNA specifically decodes AUA over AUG. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_29820.map.gz | 255.8 MB | EMDB map data format | |
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Header (meta data) | emd-29820-v30.xml emd-29820.xml | 82 KB 82 KB | Display Display | EMDB header |
Images | emd_29820.png | 236.2 KB | ||
Masks | emd_29820_msk_1.map | 512 MB | Mask map | |
Filedesc metadata | emd-29820.cif.gz | 17.3 KB | ||
Others | emd_29820_additional_1.map.gz emd_29820_half_map_1.map.gz emd_29820_half_map_2.map.gz | 422.5 MB 474.7 MB 474.7 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-29820 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-29820 | HTTPS FTP |
-Validation report
Summary document | emd_29820_validation.pdf.gz | 1.2 MB | Display | EMDB validaton report |
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Full document | emd_29820_full_validation.pdf.gz | 1.2 MB | Display | |
Data in XML | emd_29820_validation.xml.gz | 19.1 KB | Display | |
Data in CIF | emd_29820_validation.cif.gz | 22.8 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-29820 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-29820 | HTTPS FTP |
-Related structure data
Related structure data | 8g7qMC 8g7pC 8g7rC 8g7sC 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_29820.map.gz / Format: CCP4 / Size: 512 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | Ribosome bound to EF-Tu-GDPCP in the presence of paromomycin with AUG codon in the A site, A-site Ile-tRNAIle(LAU), P-site tRNAfMet and E-site tRNAIle(LAU)(Structure II) | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.86 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
File | emd_29820_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Additional map: Ribosome bound to EF-Tu-GDPCP in the presence of...
File | emd_29820_additional_1.map | ||||||||||||
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Annotation | Ribosome bound to EF-Tu-GDPCP in the presence of paromomycin with AUG codon in the A site, A-site Ile-tRNAIle(LAU), P-site tRNAfMet and E-site tRNAIle(LAU)(Structure II). Sharpened map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Ribosome bound to EF-Tu-GDPCP in the presence of...
File | emd_29820_half_map_1.map | ||||||||||||
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Annotation | Ribosome bound to EF-Tu-GDPCP in the presence of paromomycin with AUG codon in the A site, A-site Ile-tRNAIle(LAU), P-site tRNAfMet and E-site tRNAIle(LAU)(Structure II). Half map 2 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Ribosome bound to EF-Tu-GDPCP in the presence of...
File | emd_29820_half_map_2.map | ||||||||||||
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Annotation | Ribosome bound to EF-Tu-GDPCP in the presence of paromomycin with AUG codon in the A site, A-site Ile-tRNAIle(LAU), P-site tRNAfMet and E-site tRNAIle(LAU)(Structure II). Half map 1 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Structure of the Escherichia coli 70S ribosome in complex with EF...
+Supramolecule #1: Structure of the Escherichia coli 70S ribosome in complex with EF...
+Macromolecule #1: 16S Ribosomal RNA
+Macromolecule #22: Isoleucine tRNA
+Macromolecule #23: P-site initiator tRNA
+Macromolecule #24: M-M mRNA
+Macromolecule #26: 23S Ribosomal RNA
+Macromolecule #27: 5S Ribosomal RNA
+Macromolecule #2: 30S ribosomal protein S2
+Macromolecule #3: 30S ribosomal protein S3
+Macromolecule #4: 30S ribosomal protein S4
+Macromolecule #5: 30S ribosomal protein S5
+Macromolecule #6: 30S ribosomal protein S6
+Macromolecule #7: 30S ribosomal protein S7
+Macromolecule #8: 30S ribosomal protein S8
+Macromolecule #9: 30S ribosomal protein S9
+Macromolecule #10: 30S ribosomal protein S10
+Macromolecule #11: 30S ribosomal protein S11
+Macromolecule #12: 30S ribosomal protein S12
+Macromolecule #13: 30S ribosomal protein S13
+Macromolecule #14: 30S ribosomal protein S14
+Macromolecule #15: 30S ribosomal protein S15
+Macromolecule #16: 30S ribosomal protein S16
+Macromolecule #17: 30S ribosomal protein S17
+Macromolecule #18: 30S ribosomal protein S18
+Macromolecule #19: 30S ribosomal protein S19
+Macromolecule #20: 30S ribosomal protein S20
+Macromolecule #21: 30S ribosomal protein S21
+Macromolecule #25: Elongation factor Tu
+Macromolecule #28: 50S ribosomal protein L2
+Macromolecule #29: 50S ribosomal protein L3
+Macromolecule #30: 50S ribosomal protein L4
+Macromolecule #31: 50S ribosomal protein L5
+Macromolecule #32: 50S ribosomal protein L6
+Macromolecule #33: 50S ribosomal protein L9
+Macromolecule #34: 50S ribosomal protein L11
+Macromolecule #35: 50S ribosomal protein L13
+Macromolecule #36: 50S ribosomal protein L14
+Macromolecule #37: 50S ribosomal protein L15
+Macromolecule #38: 50S ribosomal protein L16
+Macromolecule #39: 50S ribosomal protein L17
+Macromolecule #40: 50S ribosomal protein L18
+Macromolecule #41: 50S ribosomal protein L19
+Macromolecule #42: 50S ribosomal protein L20
+Macromolecule #43: Ribosomal protein L21
+Macromolecule #44: 50S ribosomal protein L22
+Macromolecule #45: 50S ribosomal protein L23
+Macromolecule #46: 50S ribosomal protein L24
+Macromolecule #47: 50S ribosomal protein L25
+Macromolecule #48: 50S ribosomal protein L27
+Macromolecule #49: 50S ribosomal protein L28
+Macromolecule #50: 50S ribosomal protein L29
+Macromolecule #51: 50S ribosomal protein L30
+Macromolecule #52: 50S ribosomal protein L31
+Macromolecule #53: 50S ribosomal protein L32
+Macromolecule #54: 50S ribosomal protein L33
+Macromolecule #55: 50S ribosomal protein L34
+Macromolecule #56: 50S ribosomal protein L35
+Macromolecule #57: 50S ribosomal protein L36
+Macromolecule #58: PAROMOMYCIN
+Macromolecule #59: MAGNESIUM ION
+Macromolecule #60: ISOLEUCINE
+Macromolecule #61: PHOSPHOMETHYLPHOSPHONIC ACID GUANYLATE ESTER
+Macromolecule #62: ZINC ION
+Macromolecule #63: water
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.4 |
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Grid | Model: Quantifoil R2/1 / Material: GOLD / Mesh: 200 / Support film - Material: GOLD / Support film - topology: HOLEY / Pretreatment - Type: PLASMA CLEANING / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: OTHER |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 85 % / Chamber temperature: 295 K / Instrument: LEICA EM GP |
-Electron microscopy
Microscope | TFS KRIOS |
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Image recording | Film or detector model: FEI FALCON III (4k x 4k) / Detector mode: INTEGRATING / Number grids imaged: 1 / Number real images: 10734 / Average exposure time: 1.0 sec. / Average electron dose: 40.0 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: 2.3000000000000003 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 96000 |
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
Initial model |
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Software | Name: UCSF Chimera (ver. 1.14) | ||||||
Refinement | Space: REAL / Protocol: RIGID BODY FIT | ||||||
Output model | PDB-8g7q: |