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Yorodumi- EMDB-22466: 70S ribosome stalled on long mRNA with ArfB-1 and ArfB-2 bound (+... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-22466 | |||||||||
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Title | 70S ribosome stalled on long mRNA with ArfB-1 and ArfB-2 bound (+9-III) | |||||||||
Map data | ||||||||||
Sample |
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Keywords | Ribosome / ArfB / mRNA / TRANSLATION | |||||||||
Function / homology | Function and homology information translation release factor activity / peptidyl-tRNA hydrolase / aminoacyl-tRNA hydrolase activity / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / endoribonuclease inhibitor activity ...translation release factor activity / peptidyl-tRNA hydrolase / aminoacyl-tRNA hydrolase activity / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity / positive regulation of ribosome biogenesis / negative regulation of cytoplasmic translation / four-way junction DNA binding / 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 / rescue of stalled ribosome / 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 / 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 / mRNA binding / DNA binding / RNA binding / zinc ion binding / membrane / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Escherichia coli (strain K12) (bacteria) / Escherichia coli K-12 (bacteria) / Escherichia coli (E. coli) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.3 Å | |||||||||
Authors | Carbone CE / Korostelev AA | |||||||||
Funding support | United States, 2 items
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Citation | Journal: Nat Commun / Year: 2020 Title: ArfB can displace mRNA to rescue stalled ribosomes. Authors: Christine E Carbone / Gabriel Demo / Rohini Madireddy / Egor Svidritskiy / Andrei A Korostelev / Abstract: Ribosomes stalled during translation must be rescued to replenish the pool of translation-competent ribosomal subunits. Bacterial alternative rescue factor B (ArfB) releases nascent peptides from ...Ribosomes stalled during translation must be rescued to replenish the pool of translation-competent ribosomal subunits. Bacterial alternative rescue factor B (ArfB) releases nascent peptides from ribosomes stalled on mRNAs truncated at the A site, allowing ribosome recycling. Prior structural work revealed that ArfB recognizes such ribosomes by inserting its C-terminal α-helix into the vacant mRNA tunnel. In this work, we report that ArfB can efficiently recognize a wider range of mRNA substrates, including longer mRNAs that extend beyond the A-site codon. Single-particle cryo-EM unveils that ArfB employs two modes of function depending on the mRNA length. ArfB acts as a monomer to accommodate a shorter mRNA in the ribosomal A site. By contrast, longer mRNAs are displaced from the mRNA tunnel by more than 20 Å and are stabilized in the intersubunit space by dimeric ArfB. Uncovering distinct modes of ArfB function resolves conflicting biochemical and structural studies, and may lead to re-examination of other ribosome rescue pathways, whose functions depend on mRNA lengths. | |||||||||
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_22466.map.gz | 15.8 MB | EMDB map data format | |
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Header (meta data) | emd-22466-v30.xml emd-22466.xml | 70.3 KB 70.3 KB | Display Display | EMDB header |
Images | emd_22466.png | 200.3 KB | ||
Filedesc metadata | emd-22466.cif.gz | 12.8 KB | ||
Others | emd_22466_half_map_1.map.gz emd_22466_half_map_2.map.gz | 83.8 MB 83.8 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-22466 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-22466 | HTTPS FTP |
-Validation report
Summary document | emd_22466_validation.pdf.gz | 914.4 KB | Display | EMDB validaton report |
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Full document | emd_22466_full_validation.pdf.gz | 914 KB | Display | |
Data in XML | emd_22466_validation.xml.gz | 15.3 KB | Display | |
Data in CIF | emd_22466_validation.cif.gz | 18 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-22466 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-22466 | HTTPS FTP |
-Related structure data
Related structure data | 7jt1MC 7jssC 7jswC 7jszC 7jt2C 7jt3C 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_22466.map.gz / Format: CCP4 / Size: 178 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: 1.042 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Half map: #1
File | emd_22466_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_22466_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
+Entire : 70S ribosome stalled on long mRNA with ArfB-1 and ArfB-2 bound
+Supramolecule #1: 70S ribosome stalled on long mRNA with ArfB-1 and ArfB-2 bound
+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 L13
+Macromolecule #8: 50S ribosomal protein L14
+Macromolecule #9: 50S ribosomal protein L15
+Macromolecule #10: 50S ribosomal protein L16
+Macromolecule #11: 50S ribosomal protein L17
+Macromolecule #12: 50S ribosomal protein L18
+Macromolecule #13: 50S ribosomal protein L19
+Macromolecule #14: 50S ribosomal protein L20
+Macromolecule #15: 50S ribosomal protein L21
+Macromolecule #16: 50S ribosomal protein L22
+Macromolecule #17: 50S ribosomal protein L23
+Macromolecule #18: 50S ribosomal protein L24
+Macromolecule #19: 50S ribosomal protein L25
+Macromolecule #20: 50S ribosomal protein L27
+Macromolecule #21: 50S ribosomal protein L28
+Macromolecule #22: 50S ribosomal protein L29
+Macromolecule #23: 50S ribosomal protein L30
+Macromolecule #24: 50S ribosomal protein L32
+Macromolecule #25: 50S ribosomal protein L33
+Macromolecule #26: 50S ribosomal protein L34
+Macromolecule #27: 50S ribosomal protein L35
+Macromolecule #28: 50S ribosomal protein L36
+Macromolecule #29: 30S ribosomal protein S2
+Macromolecule #30: 30S ribosomal protein S3
+Macromolecule #31: 30S ribosomal protein S4
+Macromolecule #32: 30S ribosomal protein S5
+Macromolecule #33: 30S ribosomal protein S6
+Macromolecule #34: 30S ribosomal protein S7
+Macromolecule #35: 30S ribosomal protein S8
+Macromolecule #36: 30S ribosomal protein S9
+Macromolecule #37: 30S ribosomal protein S10
+Macromolecule #38: 30S ribosomal protein S11
+Macromolecule #39: 30S ribosomal protein S12
+Macromolecule #40: 30S ribosomal protein S13
+Macromolecule #41: 30S ribosomal protein S14
+Macromolecule #42: 30S ribosomal protein S15
+Macromolecule #43: 30S ribosomal protein S16
+Macromolecule #44: 30S ribosomal protein S17
+Macromolecule #45: 30S ribosomal protein S18
+Macromolecule #46: 30S ribosomal protein S19
+Macromolecule #47: 30S ribosomal protein S20
+Macromolecule #48: 30S ribosomal protein S21
+Macromolecule #54: Peptidyl-tRNA hydrolase ArfB
+Macromolecule #49: 16S ribosomal RNA
+Macromolecule #50: 23S ribosomal RNA
+Macromolecule #51: 5S ribosomal RNA
+Macromolecule #52: tRNAfMet
+Macromolecule #53: mRNA
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7 |
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Grid | Details: unspecified |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | TFS KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 49.6 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Startup model | Type of model: OTHER / Details: Ab initio |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.3 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 14367 |
Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |