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Yorodumi- EMDB-43075: Cryo-EM structure of the Mycobacterium smegmatis 70S ribosome in ... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-43075 | ||||||||||||
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Title | Cryo-EM structure of the Mycobacterium smegmatis 70S ribosome in complex with hibernation factor Rv2629 (Balon) (Structure 5) | ||||||||||||
Map data | Mycobacterium smegmatis 70S ribosome in complex with hibernation factor Rv2629 (Balon). Map | ||||||||||||
Sample |
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Keywords | cryo-EM / mycobacteria / hibernation / Rv2629 / Balon / RIBOSOME | ||||||||||||
Function / homology | Function and homology information peptidoglycan-based cell wall / small ribosomal subunit rRNA binding / large ribosomal subunit / regulation of translation / small ribosomal subunit / 5S rRNA binding / large ribosomal subunit rRNA binding / transferase activity / ribosomal large subunit assembly / cytosolic large ribosomal subunit ...peptidoglycan-based cell wall / small ribosomal subunit rRNA binding / large ribosomal subunit / regulation of translation / small ribosomal subunit / 5S rRNA binding / large ribosomal subunit rRNA binding / transferase activity / ribosomal large subunit assembly / cytosolic large ribosomal subunit / tRNA binding / response to hypoxia / rRNA binding / ribosome / structural constituent of ribosome / ribonucleoprotein complex / translation / mRNA binding / zinc ion binding / metal ion binding / plasma membrane / cytosol / cytoplasm Similarity search - Function | ||||||||||||
Biological species | Mycolicibacterium smegmatis MC2 155 (bacteria) / Mycobacterium tuberculosis H37Rv (bacteria) | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.1 Å | ||||||||||||
Authors | Rybak MY / Helena-Bueno K / Hill CH / Melnikov SV / Gagnon MG | ||||||||||||
Funding support | United States, 3 items
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Citation | Journal: Nature / Year: 2024 Title: A new family of bacterial ribosome hibernation factors. Authors: Karla Helena-Bueno / Mariia Yu Rybak / Chinenye L Ekemezie / Rudi Sullivan / Charlotte R Brown / Charlotte Dingwall / Arnaud Baslé / Claudia Schneider / James P R Connolly / James N Blaza / ...Authors: Karla Helena-Bueno / Mariia Yu Rybak / Chinenye L Ekemezie / Rudi Sullivan / Charlotte R Brown / Charlotte Dingwall / Arnaud Baslé / Claudia Schneider / James P R Connolly / James N Blaza / Bálint Csörgő / Patrick J Moynihan / Matthieu G Gagnon / Chris H Hill / Sergey V Melnikov / Abstract: To conserve energy during starvation and stress, many organisms use hibernation factor proteins to inhibit protein synthesis and protect their ribosomes from damage. In bacteria, two families of ...To conserve energy during starvation and stress, many organisms use hibernation factor proteins to inhibit protein synthesis and protect their ribosomes from damage. In bacteria, two families of hibernation factors have been described, but the low conservation of these proteins and the huge diversity of species, habitats and environmental stressors have confounded their discovery. Here, by combining cryogenic electron microscopy, genetics and biochemistry, we identify Balon, a new hibernation factor in the cold-adapted bacterium Psychrobacter urativorans. We show that Balon is a distant homologue of the archaeo-eukaryotic translation factor aeRF1 and is found in 20% of representative bacteria. During cold shock or stationary phase, Balon occupies the ribosomal A site in both vacant and actively translating ribosomes in complex with EF-Tu, highlighting an unexpected role for EF-Tu in the cellular stress response. Unlike typical A-site substrates, Balon binds to ribosomes in an mRNA-independent manner, initiating a new mode of ribosome hibernation that can commence while ribosomes are still engaged in protein synthesis. Our work suggests that Balon-EF-Tu-regulated ribosome hibernation is a ubiquitous bacterial stress-response mechanism, and we demonstrate that putative Balon homologues in Mycobacteria bind to ribosomes in a similar fashion. This finding calls for a revision of the current model of ribosome hibernation inferred from common model organisms and holds numerous implications for how we understand and study ribosome hibernation. | ||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_43075.map.gz | 255.3 MB | EMDB map data format | |
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Header (meta data) | emd-43075-v30.xml emd-43075.xml | 84.7 KB 84.7 KB | Display Display | EMDB header |
Images | emd_43075.png | 150.5 KB | ||
Masks | emd_43075_msk_1.map | 512 MB | Mask map | |
Filedesc metadata | emd-43075.cif.gz | 15.8 KB | ||
Others | emd_43075_additional_1.map.gz emd_43075_half_map_1.map.gz emd_43075_half_map_2.map.gz | 424 MB 476 MB 476 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-43075 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-43075 | HTTPS FTP |
-Validation report
Summary document | emd_43075_validation.pdf.gz | 1.1 MB | Display | EMDB validaton report |
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Full document | emd_43075_full_validation.pdf.gz | 1.1 MB | Display | |
Data in XML | emd_43075_validation.xml.gz | 19.2 KB | Display | |
Data in CIF | emd_43075_validation.cif.gz | 22.7 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-43075 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-43075 | HTTPS FTP |
-Related structure data
Related structure data | 8v9kMC 8rd8C 8rdvC 8rdwC 8v9jC 8v9lC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_43075.map.gz / Format: CCP4 / Size: 512 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | Mycobacterium smegmatis 70S ribosome in complex with hibernation factor Rv2629 (Balon). Map | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.85 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
File | emd_43075_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Additional map: Mycobacterium smegmatis 70S ribosome in complex with hibernation...
File | emd_43075_additional_1.map | ||||||||||||
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Annotation | Mycobacterium smegmatis 70S ribosome in complex with hibernation factor Rv2629 (Balon). Sharpened map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Mycobacterium smegmatis 70S ribosome in complex with hibernation...
File | emd_43075_half_map_1.map | ||||||||||||
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Annotation | Mycobacterium smegmatis 70S ribosome in complex with hibernation factor Rv2629 (Balon). Half-map 2 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Mycobacterium smegmatis 70S ribosome in complex with hibernation...
File | emd_43075_half_map_2.map | ||||||||||||
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Annotation | Mycobacterium smegmatis 70S ribosome in complex with hibernation factor Rv2629 (Balon). Half-map 1 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Cryo-EM structure of the Mycobacterium smegmatis 70S ribosome in ...
+Supramolecule #1: Cryo-EM structure of the Mycobacterium smegmatis 70S ribosome in ...
+Macromolecule #1: 16S Ribosomal RNA
+Macromolecule #22: poly-U mRNA
+Macromolecule #23: pe/E deacylated phenylanaline-tRNA
+Macromolecule #25: 23S Ribosomal RNA
+Macromolecule #26: 5S Ribosomal RNA
+Macromolecule #2: 30S Ribosomal Protein S2
+Macromolecule #3: 30S ribosomal protein S3
+Macromolecule #4: 30S ribosomal protein S4
+Macromolecule #5: Small ribosomal subunit protein uS5
+Macromolecule #6: 30S ribosomal protein S6
+Macromolecule #7: 30S ribosomal protein S7
+Macromolecule #8: Small ribosomal subunit protein uS8
+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: Small ribosomal subunit protein uS14B
+Macromolecule #15: Small ribosomal subunit protein uS15
+Macromolecule #16: 30S ribosomal protein S16
+Macromolecule #17: 30S ribosomal protein S17
+Macromolecule #18: Small ribosomal subunit protein bS18B
+Macromolecule #19: 30S ribosomal protein S19
+Macromolecule #20: 30S ribosomal protein S20
+Macromolecule #21: 30S Ribosomal Protein S22
+Macromolecule #24: Ribosome hibernation factor Balon (Rv2629)
+Macromolecule #27: 50S ribosomal protein L2
+Macromolecule #28: 50S ribosomal protein L3
+Macromolecule #29: Large ribosomal subunit protein uL4
+Macromolecule #30: Large ribosomal subunit protein uL5
+Macromolecule #31: 50S ribosomal protein L6
+Macromolecule #32: 50S ribosomal protein L9
+Macromolecule #33: 50S ribosomal protein L10
+Macromolecule #34: 50S ribosomal protein L11
+Macromolecule #35: Large ribosomal subunit protein uL1
+Macromolecule #36: Large ribosomal subunit protein uL13
+Macromolecule #37: 50S ribosomal protein L14
+Macromolecule #38: 50S ribosomal protein L15
+Macromolecule #39: Large ribosomal subunit protein uL16
+Macromolecule #40: 50S ribosomal protein L17
+Macromolecule #41: Large ribosomal subunit protein uL18
+Macromolecule #42: 50S ribosomal protein L19
+Macromolecule #43: Large ribosomal subunit protein bL20
+Macromolecule #44: Large ribosomal subunit protein bL21
+Macromolecule #45: Large ribosomal subunit protein uL22
+Macromolecule #46: Large ribosomal subunit protein uL23
+Macromolecule #47: 50S ribosomal protein L24
+Macromolecule #48: 50S ribosomal protein L25
+Macromolecule #49: 50S ribosomal protein L27
+Macromolecule #50: Large ribosomal subunit protein bL28
+Macromolecule #51: 50S ribosomal protein L29
+Macromolecule #52: 50S ribosomal protein L30
+Macromolecule #53: Large ribosomal subunit protein bL31
+Macromolecule #54: 50S ribosomal protein L32
+Macromolecule #55: Large ribosomal subunit protein bL33A
+Macromolecule #56: 50S ribosomal protein L34
+Macromolecule #57: 50S ribosomal protein L35
+Macromolecule #58: 50S ribosomal protein L36
+Macromolecule #59: 50S Ribosomal Protein L37
+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 Details: 20 mM HEPES-KOH, 60 mM KCl, 10 mM MgCl2, 1 mM dithiothreitol |
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Grid | Model: Quantifoil R2/1 / Material: GOLD / Mesh: 200 / Support film - Material: GOLD / Support film - topology: HOLEY / Pretreatment - Type: PLASMA CLEANING / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: OTHER |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 85 % / Chamber temperature: 295 K / Instrument: LEICA EM GP |
-Electron microscopy
Microscope | TFS KRIOS |
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Image recording | Film or detector model: FEI FALCON III (4k x 4k) / Detector mode: INTEGRATING / Number grids imaged: 1 / Number real images: 9546 / Average electron dose: 40.18 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 100.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.3000000000000003 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 96000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |