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Open data
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Basic information
| Entry | Database: EMDB / ID: EMD-10905 | |||||||||
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| Title | Structure of the P+9 stalled ribosome complex | |||||||||
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Keywords | translation / ribosome / rescue / release | |||||||||
| Function / homology | Function and homology informationstringent response / transcriptional attenuation / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity / positive regulation of ribosome biogenesis / translational termination / negative regulation of cytoplasmic translation / DnaA-L2 complex / translation repressor activity / negative regulation of DNA-templated DNA replication initiation ...stringent response / transcriptional attenuation / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity / positive regulation of ribosome biogenesis / translational termination / negative regulation of cytoplasmic translation / DnaA-L2 complex / translation repressor activity / negative regulation of DNA-templated DNA replication initiation / mRNA regulatory element binding translation repressor activity / assembly of large subunit precursor of preribosome / ribosome assembly / cytosolic ribosome assembly / response to reactive oxygen species / regulation of cell growth / DNA-templated transcription termination / response to radiation / mRNA 5'-UTR binding / ribosome biogenesis / large ribosomal subunit / ribosome binding / transferase activity / ribosomal small subunit biogenesis / ribosomal small subunit assembly / small ribosomal subunit / small ribosomal subunit rRNA binding / 5S 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 / ribonucleoprotein complex / response to antibiotic / negative regulation of DNA-templated transcription / mRNA binding / DNA binding / RNA binding / zinc ion binding / metal ion binding / cytosol / cytoplasm Similarity search - Function | |||||||||
| Biological species | ![]() | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.1 Å | |||||||||
Authors | Chan K-H / Petrychenko V | |||||||||
Citation | Journal: Nat Commun / Year: 2020Title: Mechanism of ribosome rescue by alternative ribosome-rescue factor B. Authors: Kai-Hsin Chan / Valentyn Petrychenko / Claudia Mueller / Cristina Maracci / Wolf Holtkamp / Daniel N Wilson / Niels Fischer / Marina V Rodnina / ![]() Abstract: Alternative ribosome-rescue factor B (ArfB) rescues ribosomes stalled on non-stop mRNAs by releasing the nascent polypeptide from the peptidyl-tRNA. By rapid kinetics we show that ArfB selects ...Alternative ribosome-rescue factor B (ArfB) rescues ribosomes stalled on non-stop mRNAs by releasing the nascent polypeptide from the peptidyl-tRNA. By rapid kinetics we show that ArfB selects ribosomes stalled on short truncated mRNAs, rather than on longer mRNAs mimicking pausing on rare codon clusters. In combination with cryo-electron microscopy we dissect the multistep rescue pathway of ArfB, which first binds to ribosomes very rapidly regardless of the mRNA length. The selectivity for shorter mRNAs arises from the subsequent slow engagement step, as it requires longer mRNA to shift to enable ArfB binding. Engagement results in specific interactions of the ArfB C-terminal domain with the mRNA entry channel, which activates peptidyl-tRNA hydrolysis by the N-terminal domain. These data reveal how protein dynamics translate into specificity of substrate recognition and provide insights into the action of a putative rescue factor in mitochondria. | |||||||||
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Structure visualization
| Movie |
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| Structure viewer | EM map: SurfView Molmil Jmol/JSmol |
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_10905.map.gz | 466.6 MB | EMDB map data format | |
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| Header (meta data) | emd-10905-v30.xml emd-10905.xml | 81 KB 81 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_10905_fsc.xml | 10.2 KB | Display | FSC data file |
| Images | emd_10905.png | 122.8 KB | ||
| Masks | emd_10905_msk_1.map | 91.1 MB | Mask map | |
| Filedesc metadata | emd-10905.cif.gz | 15.8 KB | ||
| Others | emd_10905_half_map_1.map.gz emd_10905_half_map_2.map.gz | 71.3 MB 71.1 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-10905 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-10905 | HTTPS FTP |
-Validation report
| Summary document | emd_10905_validation.pdf.gz | 164.4 KB | Display | EMDB validaton report |
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| Full document | emd_10905_full_validation.pdf.gz | 163.9 KB | Display | |
| Data in XML | emd_10905_validation.xml.gz | 572 B | Display | |
| Data in CIF | emd_10905_validation.cif.gz | 483 B | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10905 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10905 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 6ysrMC ![]() 6yssC ![]() 6ystC ![]() 6ysuC M: atomic model generated by this map C: citing same article ( |
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| Similar structure data | |
| EM raw data | EMPIAR-10443 (Title: Mechanism of Ribosome Rescue by Alternative Release Factor BData size: 788.2 Data #1: Motion-corrected, dose-weighted micrographs & polished particles of P+9 E. coli ribosome-ArfB complex [micrographs - single frame] Data #2: Motion-corrected, dose-weighted micrographs & extracted particles of P+0 E. coli ribosome-ArfB complex [micrographs - single frame]) |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_10905.map.gz / Format: CCP4 / Size: 512 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: 0.6525 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
| File | emd_10905_msk_1.map | ||||||||||||
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-Half map: #2
| File | emd_10905_half_map_1.map | ||||||||||||
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-Half map: #1
| File | emd_10905_half_map_2.map | ||||||||||||
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Sample components
+Entire : Stalled E. Coli P+9 ribosome complex 70S-fMet-Phe-tRNAPhe-mRNA
+Supramolecule #1: Stalled E. Coli P+9 ribosome complex 70S-fMet-Phe-tRNAPhe-mRNA
+Macromolecule #1: 50S ribosomal protein L32
+Macromolecule #2: 50S ribosomal protein L33
+Macromolecule #3: 50S ribosomal protein L34
+Macromolecule #4: 50S ribosomal protein L35
+Macromolecule #5: 50S ribosomal protein L36
+Macromolecule #6: 50S ribosomal protein L10
+Macromolecule #7: 50S ribosomal protein L31
+Macromolecule #10: 50S ribosomal protein L2
+Macromolecule #11: 50S ribosomal protein L3
+Macromolecule #12: 50S ribosomal protein L4
+Macromolecule #13: 50S ribosomal protein L5
+Macromolecule #14: 50S ribosomal protein L6
+Macromolecule #15: 50S ribosomal protein L9
+Macromolecule #16: 50S ribosomal protein L11
+Macromolecule #17: 50S ribosomal protein L13
+Macromolecule #18: 50S ribosomal protein L14
+Macromolecule #19: 50S ribosomal protein L15
+Macromolecule #20: 50S ribosomal protein L16
+Macromolecule #21: 50S ribosomal protein L17
+Macromolecule #22: 50S ribosomal protein L18
+Macromolecule #23: 50S ribosomal protein L19
+Macromolecule #24: 50S ribosomal protein L20
+Macromolecule #25: 50S ribosomal protein L21
+Macromolecule #26: 50S ribosomal protein L22
+Macromolecule #27: 50S ribosomal protein L23
+Macromolecule #28: 50S ribosomal protein L24
+Macromolecule #29: 50S ribosomal protein L25
+Macromolecule #30: 50S ribosomal protein L27
+Macromolecule #31: 50S ribosomal protein L28
+Macromolecule #32: 50S ribosomal protein L29
+Macromolecule #33: 50S ribosomal protein L30
+Macromolecule #35: 30S ribosomal protein S2
+Macromolecule #36: 30S ribosomal protein S3
+Macromolecule #37: 30S ribosomal protein S4
+Macromolecule #38: 30S ribosomal protein S5
+Macromolecule #39: 30S ribosomal protein S6
+Macromolecule #40: 30S ribosomal protein S7
+Macromolecule #41: 30S ribosomal protein S8
+Macromolecule #42: 30S ribosomal protein S9
+Macromolecule #43: 30S ribosomal protein S10
+Macromolecule #44: 30S ribosomal protein S11
+Macromolecule #45: 30S ribosomal protein S12
+Macromolecule #46: 30S ribosomal protein S13
+Macromolecule #47: 30S ribosomal protein S14
+Macromolecule #48: 30S ribosomal protein S15
+Macromolecule #49: 30S ribosomal protein S16
+Macromolecule #50: 30S ribosomal protein S17
+Macromolecule #51: 30S ribosomal protein S18
+Macromolecule #52: 30S ribosomal protein S19
+Macromolecule #53: 30S ribosomal protein S20
+Macromolecule #54: 30S ribosomal protein S21
+Macromolecule #55: P-site fMet-Phe-tRNA(Phe)
+Macromolecule #8: 23S ribosomal RNA
+Macromolecule #9: 5S ribosomal RNA
+Macromolecule #34: 16S ribosomal RNA
+Macromolecule #56: P-site fMet-Phe-tRNA(Phe)
+Macromolecule #57: mRNA
+Macromolecule #58: MAGNESIUM ION
+Macromolecule #59: ZINC ION
+Macromolecule #60: CHLORIDE ION
+Macromolecule #61: SODIUM ION
+Macromolecule #62: water
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Buffer | pH: 7.4 / Details: 50 mM HEPES, 30 mM KCl, 7 mM MgCl2 |
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| Grid | Model: Quantifoil R3.5/1 / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Pretreatment - Type: GLOW DISCHARGE / Details: Custom-made glow-discharge instrument |
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 277 K / Instrument: HOMEMADE PLUNGER / Details: Manual blotting & plunge-freezing. |
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Electron microscopy
| Microscope | TFS KRIOS |
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| Specialist optics | Spherical aberration corrector: Cs image corrector (CEOS company) |
| Details | Aberration corrections performed using Cs image corrector (CEOS company) |
| Image recording | Film or detector model: FEI FALCON III (4k x 4k) / Detector mode: INTEGRATING / Digitization - Dimensions - Width: 4096 pixel / Digitization - Dimensions - Height: 4096 pixel / Number grids imaged: 1 / Average electron dose: 50.0 e/Å2 Details: Images were collected in movie mode at 40 fractions per image per second |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | C2 aperture diameter: 100.0 µm / Calibrated defocus max: 2.5 µm / Calibrated defocus min: 0.2 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 0.01 mm / Nominal defocus max: 2.5 µm / Nominal defocus min: 0.2 µm / Nominal magnification: 59000 |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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