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Open data
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Basic information
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Title | E. coli 70S-TEC complex in delivery state | ||||||||||||||||||||||||
![]() | Primary map, sharpened | ||||||||||||||||||||||||
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![]() | Ribosome / RNA polymerase / mRNA / GENE REGULATION | ||||||||||||||||||||||||
Function / homology | ![]() mRNA 5'-UTR binding / large ribosomal subunit / transferase activity / ribosomal small subunit biogenesis / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / ribosomal large subunit assembly / cytosolic small ribosomal subunit / large ribosomal subunit rRNA binding ...mRNA 5'-UTR binding / large ribosomal subunit / transferase activity / ribosomal small subunit biogenesis / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / ribosomal large subunit assembly / cytosolic small ribosomal subunit / large ribosomal subunit rRNA binding / small ribosomal subunit rRNA binding / cytosolic large ribosomal subunit / cytoplasmic translation / tRNA binding / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / translation / ribonucleoprotein complex / mRNA binding / RNA binding / cytosol / cytoplasm Similarity search - Function | ||||||||||||||||||||||||
Biological species | ![]() ![]() ![]() ![]() | ||||||||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.17 Å | ||||||||||||||||||||||||
![]() | Webster MW / Weixlbaumer A | ||||||||||||||||||||||||
Funding support | ![]() ![]()
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![]() | ![]() Title: Molecular basis of mRNA delivery to the bacterial ribosome. Authors: Michael W Webster / Adrien Chauvier / Huma Rahil / Andrea Graziadei / Kristine Charles / Nataliya Miropolskaya / Maria Takacs / Charlotte Saint-André / Juri Rappsilber / Nils G Walter / Albert Weixlbaumer / ![]() ![]() ![]() ![]() Abstract: Protein synthesis begins with the formation of a ribosome-messenger RNA (mRNA) complex. In bacteria, the small ribosomal subunit (30) is recruited to many mRNAs through base pairing with the Shine- ...Protein synthesis begins with the formation of a ribosome-messenger RNA (mRNA) complex. In bacteria, the small ribosomal subunit (30) is recruited to many mRNAs through base pairing with the Shine-Dalgarno (SD) sequence and RNA binding by ribosomal protein bS1. Translation can initiate on nascent mRNAs, and RNA polymerase (RNAP) can promote the recruitment of the pioneering 30. Here, we examined 30 recruitment to nascent mRNAs using cryo-electron microscopy, single-molecule fluorescence colocalization, and in-cell cross-linking mass spectrometry. We show that bS1 delivers the mRNA to the ribosome for SD duplex formation and 30 activation. Additionally, bS1 and RNAP stimulate translation initiation. Our work provides a mechanistic framework for how the SD duplex, ribosomal proteins, and RNAP cooperate in 30 recruitment to mRNAs and establish transcription-translation coupling. | ||||||||||||||||||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 494.1 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 78.6 KB 78.6 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 23.9 KB | Display | ![]() |
Images | ![]() | 75 KB | ||
Filedesc metadata | ![]() | 15.8 KB | ||
Others | ![]() ![]() ![]() | 33.2 MB 815.1 MB 816 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 9gr1MC ![]() 9gupC ![]() 9guqC ![]() 9gurC ![]() 9gusC ![]() 9gutC ![]() 9guuC ![]() 9guvC ![]() 9guwC ![]() 9guxC M: atomic model generated by this map C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | Primary map, sharpened | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.052 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Additional map: Additional map. Focused reconstruction of RNA polymerase from...
File | emd_51517_additional_1.map | ||||||||||||
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Annotation | Additional map. Focused reconstruction of RNA polymerase from the same set of particle images. The map has been resampled on the primary map. | ||||||||||||
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Density Histograms |
-Half map: Half map 1
File | emd_51517_half_map_1.map | ||||||||||||
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Annotation | Half map 1 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map 2
File | emd_51517_half_map_2.map | ||||||||||||
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Annotation | Half map 2 | ||||||||||||
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Density Histograms |
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Sample components
+Entire : 70S ribosome
+Supramolecule #1: 70S ribosome
+Macromolecule #1: 50S ribosomal protein L33
+Macromolecule #2: 50S ribosomal protein L34
+Macromolecule #3: 50S ribosomal protein L35
+Macromolecule #4: 50S ribosomal protein L36
+Macromolecule #6: Small ribosomal subunit protein uS2
+Macromolecule #7: Small ribosomal subunit protein uS3
+Macromolecule #8: Small ribosomal subunit protein uS4
+Macromolecule #9: Small ribosomal subunit protein uS5
+Macromolecule #10: Small ribosomal subunit protein bS6, fully modified isoform
+Macromolecule #11: Small ribosomal subunit protein uS7
+Macromolecule #12: Small ribosomal subunit protein uS8
+Macromolecule #13: Small ribosomal subunit protein uS9
+Macromolecule #14: Small ribosomal subunit protein uS10
+Macromolecule #15: Small ribosomal subunit protein uS11
+Macromolecule #16: Small ribosomal subunit protein uS12
+Macromolecule #17: Small ribosomal subunit protein uS13
+Macromolecule #18: Small ribosomal subunit protein uS14
+Macromolecule #19: Small ribosomal subunit protein uS15
+Macromolecule #20: 30S ribosomal protein S16
+Macromolecule #21: Small ribosomal subunit protein uS17
+Macromolecule #22: Small ribosomal subunit protein bS18
+Macromolecule #23: 30S ribosomal protein S19
+Macromolecule #24: Small ribosomal subunit protein bS20
+Macromolecule #25: Small ribosomal subunit protein bS21
+Macromolecule #30: 50S ribosomal protein L2
+Macromolecule #31: Large ribosomal subunit protein uL3
+Macromolecule #32: 50S ribosomal protein L4
+Macromolecule #33: 50S ribosomal protein L5
+Macromolecule #34: 50S ribosomal protein L6
+Macromolecule #35: 50S ribosomal protein L9
+Macromolecule #36: 50S ribosomal protein L13
+Macromolecule #37: 50S ribosomal protein L14
+Macromolecule #38: 50S ribosomal protein L15
+Macromolecule #39: 50S ribosomal protein L16
+Macromolecule #40: 50S ribosomal protein L17
+Macromolecule #41: 50S ribosomal protein L18
+Macromolecule #42: 50S ribosomal protein L19
+Macromolecule #43: 50S ribosomal protein L20
+Macromolecule #44: 50S ribosomal protein L21
+Macromolecule #45: 50S ribosomal protein L22
+Macromolecule #46: 50S ribosomal protein L23
+Macromolecule #47: 50S ribosomal protein L24
+Macromolecule #48: 50S ribosomal protein L25
+Macromolecule #49: 50S ribosomal protein L27
+Macromolecule #50: 50S ribosomal protein L28
+Macromolecule #51: 50S ribosomal protein L29
+Macromolecule #52: 50S ribosomal protein L30
+Macromolecule #53: 50S ribosomal protein L32
+Macromolecule #5: 16S ribosomal RNA
+Macromolecule #26: mRNA in ribosome channel
+Macromolecule #27: Phe-NH-tRNA(Phe) A-site
+Macromolecule #28: 23S ribosomal RNA
+Macromolecule #29: 5S ribosomal RNA
+Macromolecule #54: mRNA in SD-aSD duplex
+Macromolecule #55: tRNA(fmet) P-site
+Macromolecule #56: ZINC ION
+Macromolecule #57: POTASSIUM ION
+Macromolecule #58: MAGNESIUM ION
+Macromolecule #59: PHENYLALANINE
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 8 |
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Grid | Model: Quantifoil R2/2 / Material: COPPER / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: AIR |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 283 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | TFS KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 40.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.5 µm / Nominal defocus min: 0.7000000000000001 µm |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |