+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-12857 | |||||||||
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Title | 30S ribosomal subunit from E. coli | |||||||||
Map data | ||||||||||
Sample |
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Keywords | ribosome / 30S subunit / RNA | |||||||||
Function / homology | Function and homology information mRNA 5'-UTR binding / small ribosomal subunit rRNA binding / ribosomal small subunit assembly / small ribosomal subunit / cytosolic small ribosomal subunit / cytoplasmic translation / tRNA binding / rRNA binding / ribosome / structural constituent of ribosome ...mRNA 5'-UTR binding / small ribosomal subunit rRNA binding / ribosomal small subunit assembly / small ribosomal subunit / cytosolic small ribosomal subunit / cytoplasmic translation / tRNA binding / rRNA binding / ribosome / structural constituent of ribosome / translation / mRNA binding / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Escherichia coli str. K-12 substr. MG1655 (bacteria) / Escherichia coli (strain K12) (bacteria) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.05 Å | |||||||||
Authors | Maksimova E / Korepanov A | |||||||||
Funding support | Russian Federation, 1 items
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Citation | Journal: Int J Mol Sci / Year: 2021 Title: RbfA Is Involved in Two Important Stages of 30S Subunit Assembly: Formation of the Central Pseudoknot and Docking of Helix 44 to the Decoding Center. Authors: Elena M Maksimova / Alexey P Korepanov / Olesya V Kravchenko / Timur N Baymukhametov / Alexander G Myasnikov / Konstantin S Vassilenko / Zhanna A Afonina / Elena A Stolboushkina / Abstract: Ribosome biogenesis is a highly coordinated and complex process that requires numerous assembly factors that ensure prompt and flawless maturation of ribosomal subunits. Despite the increasing amount ...Ribosome biogenesis is a highly coordinated and complex process that requires numerous assembly factors that ensure prompt and flawless maturation of ribosomal subunits. Despite the increasing amount of data collected, the exact role of most assembly factors and mechanistic details of their operation remain unclear, mainly due to the shortage of high-resolution structural information. Here, using cryo-electron microscopy, we characterized 30S ribosomal particles isolated from an strain with a deleted gene for the RbfA factor. The cryo-EM maps for pre-30S subunits were divided into six classes corresponding to consecutive assembly intermediates: from the particles with a completely unresolved head domain and unfolded central pseudoknot to almost mature 30S subunits with well-resolved body, platform, and head domains and partially distorted helix 44. The structures of two predominant 30S intermediates belonging to most populated classes obtained at 2.7 Å resolutions indicate that RbfA acts at two distinctive 30S assembly stages: early formation of the central pseudoknot including folding of the head, and positioning of helix 44 in the decoding center at a later stage. Additionally, it was shown that the formation of the central pseudoknot may promote stabilization of the head domain, likely through the RbfA-dependent maturation of the neck helix 28. An update to the model of factor-dependent 30S maturation is proposed, suggesting that RfbA is involved in most of the subunit assembly process. | |||||||||
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_12857.map.gz | 13.6 MB | EMDB map data format | |
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Header (meta data) | emd-12857-v30.xml emd-12857.xml | 50.1 KB 50.1 KB | Display Display | EMDB header |
Images | emd_12857.png | 137.1 KB | ||
Filedesc metadata | emd-12857.cif.gz | 8.9 KB | ||
Others | emd_12857_additional_1.map.gz emd_12857_additional_2.map.gz emd_12857_additional_3.map.gz emd_12857_additional_4.map.gz emd_12857_half_map_1.map.gz emd_12857_half_map_2.map.gz | 6 MB 8.4 MB 226.2 MB 226.2 MB 226.7 MB 226.7 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-12857 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-12857 | HTTPS FTP |
-Validation report
Summary document | emd_12857_validation.pdf.gz | 827.5 KB | Display | EMDB validaton report |
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Full document | emd_12857_full_validation.pdf.gz | 827.1 KB | Display | |
Data in XML | emd_12857_validation.xml.gz | 15.8 KB | Display | |
Data in CIF | emd_12857_validation.cif.gz | 18.6 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12857 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12857 | HTTPS FTP |
-Related structure data
Related structure data | 7oe1MC 7oe0C 7oi0C 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_12857.map.gz / Format: CCP4 / Size: 244.1 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: 0.86 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Additional map: 30S head
File | emd_12857_additional_1.map | ||||||||||||
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Annotation | 30S head | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: 30S body
File | emd_12857_additional_2.map | ||||||||||||
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Annotation | 30S body | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: 30S head half A map
File | emd_12857_additional_3.map | ||||||||||||
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Annotation | 30S head half A map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: 30S head half B map
File | emd_12857_additional_4.map | ||||||||||||
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Annotation | 30S head half B map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: 30S body
File | emd_12857_half_map_1.map | ||||||||||||
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Annotation | 30S body | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: 30S body
File | emd_12857_half_map_2.map | ||||||||||||
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Annotation | 30S body | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : 30S ribosomal subunit from E.coli
+Supramolecule #1: 30S ribosomal subunit from E.coli
+Macromolecule #1: 16S rRNA
+Macromolecule #2: 30S ribosomal protein S4
+Macromolecule #3: 30S ribosomal protein S5
+Macromolecule #4: 30S ribosomal protein S6, fully modified isoform
+Macromolecule #5: 30S ribosomal protein S8
+Macromolecule #6: 30S ribosomal protein S11
+Macromolecule #7: 30S ribosomal protein S12
+Macromolecule #8: 30S ribosomal protein S15
+Macromolecule #9: 30S ribosomal protein S16
+Macromolecule #10: 30S ribosomal protein S17
+Macromolecule #11: 30S ribosomal protein S18
+Macromolecule #12: 30S ribosomal protein S20
+Macromolecule #13: 30S ribosomal protein S2
+Macromolecule #14: 30S ribosomal protein S21
+Macromolecule #15: 30S ribosomal protein S3
+Macromolecule #16: 30S ribosomal protein S7
+Macromolecule #17: 30S ribosomal protein S9
+Macromolecule #18: 30S ribosomal protein S10
+Macromolecule #19: 30S ribosomal protein S13
+Macromolecule #20: 30S ribosomal protein S14
+Macromolecule #21: 30S ribosomal protein S19
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 |
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Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: FEI FALCON II (4k x 4k) / Average electron dose: 2.0 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: EMDB MAP |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.05 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 169371 |
Initial angle assignment | Type: PROJECTION MATCHING |
Final angle assignment | Type: PROJECTION MATCHING |
-Atomic model buiding 1
Initial model |
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Refinement | Space: REAL / Protocol: RIGID BODY FIT | ||||||||||||||||||||||||||||||||||||||||||||
Output model | PDB-7oe1: |