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Yorodumi- EMDB-66122: Structure of E.coli ribosome in complex with an engineered arrest... -
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Open data
- Basic information
Basic information
| Entry |  | |||||||||
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| Title | Structure of E.coli ribosome in complex with an engineered arrest peptide and trigger factor | |||||||||
|  Map data | Map | |||||||||
|  Sample | 
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|  Keywords | Ribosomal nascent chain / ribosomal arrest peptide / ribosomal translation / ribosome stalling / 70S Ribosome / RIBOSOME | |||||||||
| Function / homology |  Function and homology information 'de novo' cotranslational protein folding / stress response to copper ion / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / protein unfolding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / endoribonuclease inhibitor activity ...'de novo' cotranslational protein folding / stress response to copper ion / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / protein unfolding / 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 / protein folding chaperone / 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 / peptidylprolyl isomerase / peptidyl-prolyl cis-trans isomerase activity / regulation of cell growth / DNA-templated transcription termination / response to radiation / maintenance of translational fidelity / mRNA 5'-UTR binding / protein transport / ribosome biogenesis / large ribosomal subunit / regulation of translation / ribosome binding / transferase activity / response to heat / 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 / cell division / response to antibiotic / negative regulation of DNA-templated transcription / mRNA binding / DNA binding / RNA binding / zinc ion binding / identical protein binding / membrane / cytoplasm / cytosol Similarity search - Function | |||||||||
| Biological species |   Escherichia coli (E. coli) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 2.76 Å | |||||||||
|  Authors | Sriramoju MK / Ko TP / Draczkowski P / Hsu STD | |||||||||
| Funding support |  Taiwan, 2 items 
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|  Citation |  Journal: Nucleic Acids Res / Year: 2025 Title: Structural basis of enhanced stalling efficiency of an engineered ribosome arrest peptide. Authors: Manoj Kumar Sriramoju / Tzu-Ping Ko / Piotr Draczkowski / Shang-Te Danny Hsu /    Abstract: Ribosome stalling is a critical regulatory mechanism in protein synthesis, controlling the rate and fidelity of translation. Arrest peptides, short sequences within nascent chains, can induce ...Ribosome stalling is a critical regulatory mechanism in protein synthesis, controlling the rate and fidelity of translation. Arrest peptides, short sequences within nascent chains, can induce ribosome stalling, providing insights into the dynamics of translation and potential therapeutic targets. In this study, we investigated the molecular mechanisms of ribosome stalling induced by an engineered ribosomal arrest peptide (eRAP). We used cryo-electron microscopy and biochemical assays to characterize the interactions between eRAP, the ribosome, and accessory factors such as the trigger factor. Our results reveal intricate details of the eRAP-induced ribosome stalling, including the conformational changes in the ribosomal tunnel and the nascent chain. We also observed interactions between eRAP and specific ribosomal components, highlighting the role of key amino acids in mediating ribosome stalling. Furthermore, comparison with other stalling mechanisms, such as those induced by antibiotics or natural nascent peptides, elucidates the unique features of eRAP-induced stalling. Overall, our findings provide a comprehensive understanding of ribosome stalling by arrest peptides, shedding light on the fundamental processes of translation and offering potential avenues for therapeutic interventions targeting translation regulation. | |||||||||
| History | 
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- Structure visualization
Structure visualization
| Supplemental images | 
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- Downloads & links
Downloads & links
-EMDB archive
| Map data |  emd_66122.map.gz | 456.8 MB |  EMDB map data format | |
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| Header (meta data) |  emd-66122-v30.xml  emd-66122.xml | 84.8 KB 84.8 KB | Display Display |  EMDB header | 
| FSC (resolution estimation) |  emd_66122_fsc.xml | 20.4 KB | Display |  FSC data file | 
| Images |  emd_66122.png | 144.3 KB | ||
| Filedesc metadata |  emd-66122.cif.gz | 15.4 KB | ||
| Others |  emd_66122_half_map_1.map.gz  emd_66122_half_map_2.map.gz | 843.2 MB 843.2 MB | ||
| Archive directory |  http://ftp.pdbj.org/pub/emdb/structures/EMD-66122  ftp://ftp.pdbj.org/pub/emdb/structures/EMD-66122 | HTTPS FTP | 
-Validation report
| Summary document |  emd_66122_validation.pdf.gz | 993.2 KB | Display |  EMDB validaton report | 
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| Full document |  emd_66122_full_validation.pdf.gz | 992.8 KB | Display | |
| Data in XML |  emd_66122_validation.xml.gz | 30.4 KB | Display | |
| Data in CIF |  emd_66122_validation.cif.gz | 40.4 KB | Display | |
| Arichive directory |  https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-66122  ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-66122 | HTTPS FTP | 
-Related structure data
| Related structure data |  9wnrMC  9wnqC 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_66122.map.gz / Format: CCP4 / Size: 909.2 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | Map | ||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
 
 Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.1 Å | ||||||||||||||||||||||||||||||||||||
| Density | 
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML: 
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-Supplemental data
-Half map: HalfMap-B
| File | emd_66122_half_map_1.map | ||||||||||||
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| Annotation | HalfMap-B | ||||||||||||
| Projections & Slices | 
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| Density Histograms | 
-Half map: HalfMap-A
| File | emd_66122_half_map_2.map | ||||||||||||
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| Annotation | HalfMap-A | ||||||||||||
| Projections & Slices | 
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| Density Histograms | 
- Sample components
Sample components
+Entire : E.coli 70S Ribosome in complex with eRAP
+Supramolecule #1: E.coli 70S Ribosome in complex with eRAP
+Macromolecule #1: Large ribosomal subunit protein bL28
+Macromolecule #2: Large ribosomal subunit protein uL29
+Macromolecule #3: Large ribosomal subunit protein uL30
+Macromolecule #4: Large ribosomal subunit protein bL32
+Macromolecule #5: Large ribosomal subunit protein bL33
+Macromolecule #6: 50S ribosomal protein L34
+Macromolecule #7: Large ribosomal subunit protein bL35
+Macromolecule #8: 50S ribosomal protein L36
+Macromolecule #10: 30S ribosomal protein S2
+Macromolecule #11: 30S ribosomal protein S3
+Macromolecule #12: 30S ribosomal protein S4
+Macromolecule #13: 30S ribosomal protein S5
+Macromolecule #14: 30S ribosomal protein S6, fully modified isoform
+Macromolecule #15: 30S ribosomal protein S7
+Macromolecule #16: 30S ribosomal protein S8
+Macromolecule #17: 30S ribosomal protein S9
+Macromolecule #18: 30S ribosomal protein S10
+Macromolecule #19: Small ribosomal subunit protein uS11
+Macromolecule #20: Small ribosomal subunit protein uS12
+Macromolecule #21: 30S ribosomal protein S13
+Macromolecule #22: 30S ribosomal protein S14
+Macromolecule #23: 30S ribosomal protein S15
+Macromolecule #24: 30S ribosomal protein S16
+Macromolecule #25: 30S ribosomal protein S17
+Macromolecule #26: 30S ribosomal protein S18
+Macromolecule #27: 30S ribosomal protein S19
+Macromolecule #28: 30S ribosomal protein S20
+Macromolecule #29: 30S ribosomal protein S21
+Macromolecule #30: Trigger factor
+Macromolecule #35: Large ribosomal subunit protein uL2
+Macromolecule #36: 50S ribosomal protein L3
+Macromolecule #37: 50S ribosomal protein L4
+Macromolecule #38: Large ribosomal subunit protein uL5
+Macromolecule #39: Large ribosomal subunit protein uL6
+Macromolecule #40: 50S ribosomal protein L9
+Macromolecule #41: 50S ribosomal protein L13
+Macromolecule #42: 50S ribosomal protein L14
+Macromolecule #43: 50S ribosomal protein L15
+Macromolecule #44: 50S ribosomal protein L16
+Macromolecule #45: Large ribosomal subunit protein bL17
+Macromolecule #46: Large ribosomal subunit protein uL18
+Macromolecule #47: Large ribosomal subunit protein bL19
+Macromolecule #48: Large ribosomal subunit protein bL20
+Macromolecule #49: 50S ribosomal protein L21
+Macromolecule #50: 50S ribosomal protein L22
+Macromolecule #51: Large ribosomal subunit protein uL23
+Macromolecule #52: 50S ribosomal protein L24
+Macromolecule #53: 50S ribosomal protein L25
+Macromolecule #55: Large ribosomal subunit protein bL27
+Macromolecule #56: eRAP-NC
+Macromolecule #9: 16S rRNA
+Macromolecule #31: E-tRNA
+Macromolecule #32: P-tRNA(PRO), A-tRNA(PRO)
+Macromolecule #33: 5S rRNA
+Macromolecule #34: 23S rRNA
+Macromolecule #54: mRNA
+Macromolecule #57: ZINC ION
+Macromolecule #58: PROLINE
-Experimental details
-Structure determination
| Method | cryo EM | 
|---|---|
|  Processing | single particle reconstruction | 
| Aggregation state | particle | 
- Sample preparation
Sample preparation
| Concentration | 0.42 mg/mL | 
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| Buffer | pH: 7.6 Details: 20 mM HEPES (pH 7.6), 140 mM NH4Cl, 25 mM MgCl2, 2 mM DTT | 
| Grid | Model: Quantifoil / Support film - Material: CARBON / Support film - topology: CONTINUOUS | 
| Vitrification | Cryogen name: NITROGEN / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV | 
- Electron microscopy
Electron microscopy
| Microscope | TFS KRIOS | 
|---|---|
| Image recording | Film or detector model: GATAN K3 (6k x 4k) / Number real images: 15280 / Average exposure time: 2.5 sec. / Average electron dose: 38.0 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: 1.8 µm / Nominal defocus min: 1.0 µm | 
| Sample stage | Cooling holder cryogen: NITROGEN | 
| Experimental equipment |  Model: Titan Krios / Image courtesy: FEI Company | 
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