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Yorodumi- EMDB-11713: Structure of accomodated trans-translation complex on E. Coli sta... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-11713 | |||||||||||||||
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Title | Structure of accomodated trans-translation complex on E. Coli stalled ribosome. | |||||||||||||||
Map data | Sharpened not masked map | |||||||||||||||
Sample |
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Keywords | Trans-translation / tmRNA / SmpB / Ribosome / TRANSLATION | |||||||||||||||
Function / homology | Function and homology information trans-translation / 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 / RNA-binding transcription regulator activity ...trans-translation / 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 / 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 translational initiation / negative regulation of DNA-templated DNA replication initiation / regulation of mRNA stability / mRNA regulatory element binding translation repressor activity / 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 / translational initiation / 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 / large ribosomal subunit rRNA binding / small ribosomal subunit / 5S 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 (E. coli) / Escherichia coli K-12 (bacteria) | |||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.08 Å | |||||||||||||||
Authors | Guyomar C / D'Urso G | |||||||||||||||
Funding support | France, 4 items
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Citation | Journal: Nat Commun / Year: 2021 Title: Structures of tmRNA and SmpB as they transit through the ribosome. Authors: Charlotte Guyomar / Gaetano D'Urso / Sophie Chat / Emmanuel Giudice / Reynald Gillet / Abstract: In bacteria, trans-translation is the main rescue system, freeing ribosomes stalled on defective messenger RNAs. This mechanism is driven by small protein B (SmpB) and transfer-messenger RNA (tmRNA), ...In bacteria, trans-translation is the main rescue system, freeing ribosomes stalled on defective messenger RNAs. This mechanism is driven by small protein B (SmpB) and transfer-messenger RNA (tmRNA), a hybrid RNA known to have both a tRNA-like and an mRNA-like domain. Here we present four cryo-EM structures of the ribosome during trans-translation at resolutions from 3.0 to 3.4 Å. These include the high-resolution structure of the whole pre-accommodated state, as well as structures of the accommodated state, the translocated state, and a translocation intermediate. Together, they shed light on the movements of the tmRNA-SmpB complex in the ribosome, from its delivery by the elongation factor EF-Tu to its passage through the ribosomal A and P sites after the opening of the B1 bridges. Additionally, we describe the interactions between the tmRNA-SmpB complex and the ribosome. These explain why the process does not interfere with canonical translation. | |||||||||||||||
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_11713.map.gz | 376.9 MB | EMDB map data format | |
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Header (meta data) | emd-11713-v30.xml emd-11713.xml | 88.5 KB 88.5 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_11713_fsc.xml | 17 KB | Display | FSC data file |
Images | emd_11713.png | 178.6 KB | ||
Masks | emd_11713_msk_1.map | 421.9 MB | Mask map | |
Filedesc metadata | emd-11713.cif.gz | 15.6 KB | ||
Others | emd_11713_additional_1.map.gz emd_11713_half_map_1.map.gz emd_11713_half_map_2.map.gz | 336.9 MB 337.9 MB 338.5 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-11713 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-11713 | HTTPS FTP |
-Validation report
Summary document | emd_11713_validation.pdf.gz | 1.2 MB | Display | EMDB validaton report |
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Full document | emd_11713_full_validation.pdf.gz | 1.2 MB | Display | |
Data in XML | emd_11713_validation.xml.gz | 24.1 KB | Display | |
Data in CIF | emd_11713_validation.cif.gz | 31.5 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-11713 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-11713 | HTTPS FTP |
-Related structure data
Related structure data | 7ac7MC 7abzC 7acjC 7acrC M: atomic model generated by this map C: citing same article (ref.) |
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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_11713.map.gz / Format: CCP4 / Size: 421.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Sharpened not masked map | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.04 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_11713_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Additional map: #1
File | emd_11713_additional_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_11713_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_11713_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Structure of accomodated trans-translation complex on E. Coli sta...
+Supramolecule #1: Structure of accomodated trans-translation complex on E. Coli sta...
+Supramolecule #2: tmRNA
+Supramolecule #3: SsrA-binding protein
+Supramolecule #4: tRNA, mRNA
+Supramolecule #5: Ribosome
+Macromolecule #1: 23S ribosomal RNA
+Macromolecule #2: 16S ribosomal RNA
+Macromolecule #3: 5S ribosomal RNA
+Macromolecule #4: transfer-messenger RNA (tmRNA)
+Macromolecule #6: tRNA-Phe
+Macromolecule #7: mRNA
+Macromolecule #5: SsrA-binding protein
+Macromolecule #8: 50S ribosomal protein L27
+Macromolecule #9: 50S ribosomal protein L2
+Macromolecule #10: 50S ribosomal protein L3
+Macromolecule #11: 50S ribosomal protein L4
+Macromolecule #12: 50S ribosomal protein L5
+Macromolecule #13: 50S ribosomal protein L6
+Macromolecule #14: 50S ribosomal protein L9
+Macromolecule #15: 50S ribosomal protein L10
+Macromolecule #16: 50S ribosomal protein L13
+Macromolecule #17: 50S ribosomal protein L14
+Macromolecule #18: 50S ribosomal protein L15
+Macromolecule #19: 50S ribosomal protein L16
+Macromolecule #20: 50S ribosomal protein L17
+Macromolecule #21: 50S ribosomal protein L18
+Macromolecule #22: 50S ribosomal protein L19
+Macromolecule #23: 50S ribosomal protein L20
+Macromolecule #24: 50S ribosomal protein L21
+Macromolecule #25: 50S ribosomal protein L22
+Macromolecule #26: 50S ribosomal protein L23
+Macromolecule #27: 50S ribosomal protein L24
+Macromolecule #28: 50S ribosomal protein L25
+Macromolecule #29: Nascent peptide
+Macromolecule #30: 50S ribosomal protein L28
+Macromolecule #31: 50S ribosomal protein L29
+Macromolecule #32: 50S ribosomal protein L30
+Macromolecule #33: 50S ribosomal protein L31
+Macromolecule #34: 50S ribosomal protein L32
+Macromolecule #35: 50S ribosomal protein L33
+Macromolecule #36: 50S ribosomal protein L34
+Macromolecule #37: 50S ribosomal protein L35
+Macromolecule #38: 50S ribosomal protein L36
+Macromolecule #39: 30S ribosomal protein S2
+Macromolecule #40: 30S ribosomal protein S3
+Macromolecule #41: 30S ribosomal protein S4
+Macromolecule #42: 30S ribosomal protein S5
+Macromolecule #43: 30S ribosomal protein S6
+Macromolecule #44: 30S ribosomal protein S7
+Macromolecule #45: 30S ribosomal protein S8
+Macromolecule #46: 30S ribosomal protein S9
+Macromolecule #47: 30S ribosomal protein S10
+Macromolecule #48: 30S ribosomal protein S11
+Macromolecule #49: 30S ribosomal protein S12
+Macromolecule #50: 30S ribosomal protein S13
+Macromolecule #51: 30S ribosomal protein S14
+Macromolecule #52: 30S ribosomal protein S15
+Macromolecule #53: 30S ribosomal protein S16
+Macromolecule #54: 30S ribosomal protein S17
+Macromolecule #55: 30S ribosomal protein S18
+Macromolecule #56: 30S ribosomal protein S19
+Macromolecule #57: 30S ribosomal protein S20
+Macromolecule #58: 30S ribosomal protein S21
+Macromolecule #59: MAGNESIUM ION
+Macromolecule #60: ZINC ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 Component:
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Grid | Model: Quantifoil R2/2 / Material: COPPER / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: OTHER / Details: 10mA | ||||||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 298 K / Instrument: FEI VITROBOT MARK III / Details: blot for 3.5 seconds before plunging. |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 BASE (4k x 4k) / Digitization - Dimensions - Width: 3710 pixel / Digitization - Dimensions - Height: 3838 pixel / Number real images: 22350 / Average exposure time: 10.0 sec. / Average electron dose: 29.0 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: -2.0 µm / Nominal defocus min: -0.7000000000000001 µm / Nominal magnification: 130000 |
Sample stage | Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |