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Yorodumi- EMDB-39577: E. coli 70S ribosome complexed with P. putida tRNAIle2 at the A-s... -
+ Open data
Open data
- Basic information
Basic information
| Entry |  | ||||||||||||
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| Title | E. coli 70S ribosome complexed with P. putida tRNAIle2 at the A-site and P-site | ||||||||||||
|  Map data | |||||||||||||
|  Sample | 
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|  Keywords | tRNA modification / decoding / RIBOSOME | ||||||||||||
| Function / homology |  Function and homology information negative regulation of cytoplasmic translational initiation / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / endoribonuclease inhibitor activity / positive regulation of ribosome biogenesis / RNA-binding transcription regulator activity ...negative regulation of cytoplasmic translational initiation / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / endoribonuclease inhibitor activity / positive regulation of ribosome biogenesis / RNA-binding transcription regulator activity / 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 / positive regulation of RNA splicing / 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 / mRNA binding / DNA binding / RNA binding / zinc ion binding / membrane / cytoplasm / cytosol Similarity search - Function | ||||||||||||
| Biological species |   Escherichia coli (E. coli) /  Pseudomonas putida NBRC 14164 (bacteria) | ||||||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 2.25 Å | ||||||||||||
|  Authors | Akiyama N / Ishiguro K / Shirouzu M / Suzuki T | ||||||||||||
| Funding support |  Japan, 3 items 
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|  Citation |  Journal: Nat Chem Biol / Year: 2025 Title: A tRNA modification with aminovaleramide facilitates AUA decoding in protein synthesis. Authors: Kenjyo Miyauchi / Satoshi Kimura / Naho Akiyama / Kazuki Inoue / Kensuke Ishiguro / Thien-Son Vu / Veerasak Srisuknimit / Kenta Koyama / Gosuke Hayashi / Akiko Soma / Asuteka Nagao / Mikako ...Authors: Kenjyo Miyauchi / Satoshi Kimura / Naho Akiyama / Kazuki Inoue / Kensuke Ishiguro / Thien-Son Vu / Veerasak Srisuknimit / Kenta Koyama / Gosuke Hayashi / Akiko Soma / Asuteka Nagao / Mikako Shirouzu / Akimitsu Okamoto / Matthew K Waldor / Tsutomu Suzuki /      Abstract: Modified tRNA anticodons are critical for proper mRNA translation during protein synthesis. It is generally thought that almost all bacterial tRNAs use a modified cytidine-lysidine (L)-at the first ...Modified tRNA anticodons are critical for proper mRNA translation during protein synthesis. It is generally thought that almost all bacterial tRNAs use a modified cytidine-lysidine (L)-at the first position (34) of the anticodon to decipher the AUA codon as isoleucine (Ile). Here we report that tRNAs from plant organelles and a subset of bacteria contain a new cytidine derivative, designated 2-aminovaleramididine (avaC). Like L34, avaC34 governs both Ile-charging ability and AUA decoding. Cryo-electron microscopy structural analyses revealed molecular details of codon recognition by avaC34 with a specific interaction between its terminal amide group and an mRNA residue 3'-adjacent to the AUA codon. These findings reveal the evolutionary variation of an essential tRNA modification and demonstrate the molecular basis of AUA decoding mediated by a unique tRNA modification. | ||||||||||||
| History | 
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- Structure visualization
Structure visualization
| Supplemental images | 
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- Downloads & links
Downloads & links
-EMDB archive
| Map data |  emd_39577.map.gz | 531.2 MB |  EMDB map data format | |
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| Header (meta data) |  emd-39577-v30.xml  emd-39577.xml | 87.2 KB 87.2 KB | Display Display |  EMDB header | 
| FSC (resolution estimation) |  emd_39577_fsc.xml | 18.6 KB | Display |  FSC data file | 
| Images |  emd_39577.png | 92.9 KB | ||
| Filedesc metadata |  emd-39577.cif.gz | 15.9 KB | ||
| Others |  emd_39577_additional_1.map.gz  emd_39577_half_map_1.map.gz  emd_39577_half_map_2.map.gz | 458.7 MB 461.3 MB 461.3 MB | ||
| Archive directory |  http://ftp.pdbj.org/pub/emdb/structures/EMD-39577  ftp://ftp.pdbj.org/pub/emdb/structures/EMD-39577 | HTTPS FTP | 
-Validation report
| Summary document |  emd_39577_validation.pdf.gz | 1.2 MB | Display |  EMDB validaton report | 
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| Full document |  emd_39577_full_validation.pdf.gz | 1.2 MB | Display | |
| Data in XML |  emd_39577_validation.xml.gz | 26.8 KB | Display | |
| Data in CIF |  emd_39577_validation.cif.gz | 35.9 KB | Display | |
| Arichive directory |  https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-39577  ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-39577 | HTTPS FTP | 
-Related structure data
| Related structure data |  8yuoMC  8yupC  8yuqC  8yurC  8yusC M: atomic model generated by this map C: citing same article ( | 
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| Similar structure data | Similarity search - Function & homology  F&H Search | 
- Links
Links
| EMDB pages |  EMDB (EBI/PDBe) /  EMDataResource | 
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| Related items in Molecule of the Month | 
- Map
Map
| File |  Download / File: emd_39577.map.gz / Format: CCP4 / Size: 567.9 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: 0.8285 Å | ||||||||||||||||||||||||||||||||||||
| Density | 
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML: 
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-Supplemental data
-Additional map: Before post-processing
| File | emd_39577_additional_1.map | ||||||||||||
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| Annotation | Before post-processing | ||||||||||||
| Projections & Slices | 
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| Density Histograms | 
-Half map: #2
| File | emd_39577_half_map_1.map | ||||||||||||
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| Projections & Slices | 
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| Density Histograms | 
-Half map: #1
| File | emd_39577_half_map_2.map | ||||||||||||
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| Projections & Slices | 
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| Density Histograms | 
- Sample components
Sample components
+Entire : The complex of E. coli 70S ribosome with mRNA and A-, P- site tRNA
+Supramolecule #1: The complex of E. coli 70S ribosome with mRNA and A-, P- site tRNA
+Supramolecule #2: E. coli 70S ribosome
+Supramolecule #3: mRNA
+Supramolecule #4: A-site tRNAIle2
+Macromolecule #1: 16S rRNA
+Macromolecule #22: 23S rRNA
+Macromolecule #23: 5S rRNA
+Macromolecule #53: mRNA
+Macromolecule #54: A-site tRNAIle2
+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, fully modified isoform
+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 #24: 50S ribosomal protein L2
+Macromolecule #25: 50S ribosomal protein L3
+Macromolecule #26: 50S ribosomal protein L4
+Macromolecule #27: 50S ribosomal protein L5
+Macromolecule #28: 50S ribosomal protein L6
+Macromolecule #29: 50S ribosomal protein L9
+Macromolecule #30: 50S ribosomal protein L13
+Macromolecule #31: 50S ribosomal protein L14
+Macromolecule #32: 50S ribosomal protein L15
+Macromolecule #33: 50S ribosomal protein L16
+Macromolecule #34: 50S ribosomal protein L17
+Macromolecule #35: 50S ribosomal protein L18
+Macromolecule #36: 50S ribosomal protein L19
+Macromolecule #37: 50S ribosomal protein L20
+Macromolecule #38: 50S ribosomal protein L21
+Macromolecule #39: 50S ribosomal protein L22
+Macromolecule #40: 50S ribosomal protein L23
+Macromolecule #41: 50S ribosomal protein L24
+Macromolecule #42: 50S ribosomal protein L25
+Macromolecule #43: 50S ribosomal protein L27
+Macromolecule #44: 50S ribosomal protein L28
+Macromolecule #45: 50S ribosomal protein L29
+Macromolecule #46: 50S ribosomal protein L30
+Macromolecule #47: 50S ribosomal protein L32
+Macromolecule #48: 50S ribosomal protein L33
+Macromolecule #49: 50S ribosomal protein L34
+Macromolecule #50: 50S ribosomal protein L35
+Macromolecule #51: 50S ribosomal protein L36
+Macromolecule #52: 50S ribosomal protein L31
+Macromolecule #55: MAGNESIUM ION
-Experimental details
-Structure determination
| Method | cryo EM | 
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|  Processing | single particle reconstruction | 
| Aggregation state | particle | 
- Sample preparation
Sample preparation
| Buffer | pH: 7.6 Component: 
 Details: The Buffer pH was adjusted to 7.6 using KOH. | |||||||||||||||
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| Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 | |||||||||||||||
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV | |||||||||||||||
| Details | 100nM ribosomes were incubated with 500nM tRNAs and mRNA | 
- Electron microscopy
Electron microscopy
| Microscope | FEI TITAN KRIOS | 
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| Image recording | Film or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 1 / Number real images: 6134 / Average electron dose: 50.0 e/Å2 | 
| Electron beam | Acceleration voltage: 300 kV / Electron source:  FIELD EMISSION GUN | 
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.5 µm / Nominal defocus min: 0.5 µm | 
| Experimental equipment |  Model: Titan Krios / Image courtesy: FEI Company | 
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