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Yorodumi- EMDB-41049: Cryo-EM studies of the interplay between uS2 ribosomal protein an... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-41049 | |||||||||
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Title | Cryo-EM studies of the interplay between uS2 ribosomal protein and leaderless mRNA during bacterial translation initiation | |||||||||
Map data | rpsB11-withS2-S21 | |||||||||
Sample |
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Keywords | leaderless mRNA / Cryo-EM / 70S / HflX / RIBOSOME / bS21 / uS2 | |||||||||
Function / homology | Function and homology information transcription elongation-coupled chromatin remodeling / ribosome assembly / assembly of large subunit precursor of preribosome / cytosolic ribosome assembly / regulation of DNA-templated transcription elongation / transcription antitermination / DNA-templated transcription termination / mRNA 5'-UTR binding / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding ...transcription elongation-coupled chromatin remodeling / ribosome assembly / assembly of large subunit precursor of preribosome / cytosolic ribosome assembly / regulation of DNA-templated transcription elongation / transcription antitermination / DNA-templated transcription termination / mRNA 5'-UTR binding / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / large ribosomal subunit / 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 / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / ribonucleoprotein complex / translation / response to antibiotic / mRNA binding / membrane / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Escherichia coli (E. coli) / Bacteriophage sp. (virus) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.2 Å | |||||||||
Authors | Bhattacharjee S / Gottesman ME / Frank J | |||||||||
Funding support | United States, 2 items
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Citation | Journal: J Mol Biol / Year: 2024 Title: How Dedicated Ribosomes Translate a Leaderless mRNA. Authors: Francisco J Acosta-Reyes / Sayan Bhattacharjee / Max Gottesman / Joachim Frank / Abstract: In bacteriophage λ lysogens, the λcI repressor is encoded by the leaderless transcript (lmRNA) initiated at the λpRM promoter. Translation is enhanced in rpsB mutants deficient in ribosomal ...In bacteriophage λ lysogens, the λcI repressor is encoded by the leaderless transcript (lmRNA) initiated at the λpRM promoter. Translation is enhanced in rpsB mutants deficient in ribosomal protein uS2. Although translation initiation of lmRNA is conserved in bacteria, archaea, and eukaryotes, structural insight of a lmRNA translation initiation complex is missing. Here, we use cryo-EM to solve the structures of the uS2-deficient 70S ribosome of host E. coli mutant rpsB11 and the wild-type 70S complex with λcI lmRNA and fMet-tRNA. Importantly, the uS2-deficient 70S ribosome also lacks protein bS21. The anti-Shine-Dalgarno (aSD) region is structurally supported by bS21, so that the absence of the latter causes the aSD to divert from the normal mRNA exit pathway, easing the exit of lmRNA. A π-stacking interaction between the monitor base A1493 and A(+4) of lmRNA potentially acts as a recognition signal. Coulomb charge flow, along with peristalsis-like dynamics within the mRNA entrance channel due to the increased 30S head rotation caused by the absence of uS2, are likely to facilitate the propagation of lmRNA through the ribosome. These findings lay the groundwork for future research on the mechanism of translation and the co-evolution of lmRNA and mRNA that includes the emergence of a defined ribosome-binding site of the transcript. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_41049.map.gz | 200.8 MB | EMDB map data format | |
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Header (meta data) | emd-41049-v30.xml emd-41049.xml | 75.1 KB 75.1 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_41049_fsc.xml | 14.2 KB | Display | FSC data file |
Images | emd_41049.png | 116.1 KB | ||
Masks | emd_41049_msk_1.map | 244.1 MB | Mask map | |
Filedesc metadata | emd-41049.cif.gz | 16.2 KB | ||
Others | emd_41049_half_map_1.map.gz emd_41049_half_map_2.map.gz | 226.9 MB 226.9 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-41049 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-41049 | HTTPS FTP |
-Validation report
Summary document | emd_41049_validation.pdf.gz | 1.2 MB | Display | EMDB validaton report |
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Full document | emd_41049_full_validation.pdf.gz | 1.2 MB | Display | |
Data in XML | emd_41049_validation.xml.gz | 21.5 KB | Display | |
Data in CIF | emd_41049_validation.cif.gz | 27.9 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-41049 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-41049 | HTTPS FTP |
-Related structure data
Related structure data | 8t5dMC 8t5hC 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_41049.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | rpsB11-withS2-S21 | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.83 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
File | emd_41049_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: half1
File | emd_41049_half_map_1.map | ||||||||||||
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Annotation | half1 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: half2
File | emd_41049_half_map_2.map | ||||||||||||
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Annotation | half2 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Apo 70S at 900 ms
+Supramolecule #1: Apo 70S at 900 ms
+Macromolecule #1: 50S ribosomal protein L32
+Macromolecule #2: 50S ribosomal protein L33
+Macromolecule #3: 50S ribosomal protein L34
+Macromolecule #4: 50S ribosomal protein L35
+Macromolecule #5: 50S ribosomal protein L36
+Macromolecule #8: 50S ribosomal protein L2
+Macromolecule #9: 50S ribosomal protein L3
+Macromolecule #10: 50S ribosomal protein L4
+Macromolecule #11: 50S ribosomal protein L5
+Macromolecule #12: 50S ribosomal protein L6
+Macromolecule #13: 50S ribosomal protein L13
+Macromolecule #14: 50S ribosomal protein L14
+Macromolecule #15: 50S ribosomal protein L15
+Macromolecule #16: 50S ribosomal protein L16
+Macromolecule #17: 50S ribosomal protein L17
+Macromolecule #18: 50S ribosomal protein L18
+Macromolecule #19: 50S ribosomal protein L19
+Macromolecule #20: 50S ribosomal protein L20
+Macromolecule #21: Ribosomal protein L21
+Macromolecule #22: 50S ribosomal protein L22
+Macromolecule #23: 50S ribosomal protein L23
+Macromolecule #24: 50S ribosomal protein L24
+Macromolecule #25: 50S ribosomal protein L25
+Macromolecule #26: 50S ribosomal protein L27
+Macromolecule #27: 50S ribosomal protein L28
+Macromolecule #28: 50S ribosomal protein L29
+Macromolecule #29: 50S ribosomal protein L30
+Macromolecule #30: 30S ribosomal protein S2
+Macromolecule #31: 30S ribosomal protein S3
+Macromolecule #32: 30S ribosomal protein S4
+Macromolecule #33: 30S ribosomal protein S5
+Macromolecule #34: 30S ribosomal protein S6, non-modified isoform
+Macromolecule #35: 30S ribosomal protein S7
+Macromolecule #36: 30S ribosomal protein S8
+Macromolecule #37: 30S ribosomal protein S9
+Macromolecule #38: 30S ribosomal protein S10
+Macromolecule #39: 30S ribosomal protein S11
+Macromolecule #40: 30S ribosomal protein S12
+Macromolecule #41: 30S ribosomal protein S13
+Macromolecule #42: 30S ribosomal protein S14
+Macromolecule #43: 30S ribosomal protein S15
+Macromolecule #44: 30S ribosomal protein S16
+Macromolecule #45: 30S ribosomal protein S17
+Macromolecule #46: 30S ribosomal protein S18
+Macromolecule #47: 30S ribosomal protein S19
+Macromolecule #48: 30S ribosomal protein S20
+Macromolecule #49: 30S ribosomal protein S21 (Fragment)
+Macromolecule #50: Transcription termination/antitermination protein NusG
+Macromolecule #6: 5S RNA
+Macromolecule #7: 23S RNA
+Macromolecule #51: 16s RNA
+Macromolecule #52: lmRNA
+Macromolecule #53: tRNA
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | cell |
-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: GATAN K3 (6k x 4k) / Average electron dose: 58.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: DARK FIELD / Nominal defocus max: 2.5 µm / Nominal defocus min: 1.0 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |