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Yorodumi- EMDB-51903: Late-stage 48S Initiation Complex (LS48S IC) guided by the trans-RNA -
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Open data
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Basic information
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| Title | Late-stage 48S Initiation Complex (LS48S IC) guided by the trans-RNA | ||||||||||||
Map data | main map of the mammalian late-stage 48S initiation complex | ||||||||||||
Sample |
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Keywords | 48S late-stage initiation complex / Programmable mRNA translation initiation / trans-RNA / activation of mRNA translation / TRANSLATION | ||||||||||||
| Function / homology | Function and homology informationregulation of macromolecule metabolic process / Translation initiation complex formation / Formation of the ternary complex, and subsequently, the 43S complex / Ribosomal scanning and start codon recognition / eukaryotic initiation factor eIF2 binding / Major pathway of rRNA processing in the nucleolus and cytosol / GTP hydrolysis and joining of the 60S ribosomal subunit / L13a-mediated translational silencing of Ceruloplasmin expression / SRP-dependent cotranslational protein targeting to membrane / Formation of a pool of free 40S subunits ...regulation of macromolecule metabolic process / Translation initiation complex formation / Formation of the ternary complex, and subsequently, the 43S complex / Ribosomal scanning and start codon recognition / eukaryotic initiation factor eIF2 binding / Major pathway of rRNA processing in the nucleolus and cytosol / GTP hydrolysis and joining of the 60S ribosomal subunit / L13a-mediated translational silencing of Ceruloplasmin expression / SRP-dependent cotranslational protein targeting to membrane / Formation of a pool of free 40S subunits / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / eukaryotic translation initiation factor 2 complex / formation of cytoplasmic translation initiation complex / eukaryotic 48S preinitiation complex / cellular response to chemical stress / GDP-dissociation inhibitor activity / laminin receptor activity / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / ubiquitin ligase inhibitor activity / positive regulation of signal transduction by p53 class mediator / 90S preribosome / phagocytic cup / laminin binding / endonucleolytic cleavage in ITS1 to separate SSU-rRNA from 5.8S rRNA and LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rough endoplasmic reticulum / translation regulator activity / ribosomal small subunit export from nucleus / gastrulation / MDM2/MDM4 family protein binding / translation initiation factor activity / cytosolic ribosome / class I DNA-(apurinic or apyrimidinic site) endonuclease activity / DNA-(apurinic or apyrimidinic site) lyase / GTPase activator activity / response to endoplasmic reticulum stress / positive regulation of apoptotic signaling pathway / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / small-subunit processome / spindle / cytoplasmic stress granule / rRNA processing / rhythmic process / positive regulation of canonical Wnt signaling pathway / regulation of translation / ribosome binding / virus receptor activity / ribosomal small subunit biogenesis / ribosomal small subunit assembly / small ribosomal subunit / small ribosomal subunit rRNA binding / cytosolic small ribosomal subunit / perikaryon / cytoplasmic translation / cell differentiation / tRNA binding / mitochondrial inner membrane / rRNA binding / postsynaptic density / structural constituent of ribosome / ribosome / translation / ribonucleoprotein complex / cell division / DNA repair / mRNA binding / apoptotic process / synapse / dendrite / centrosome / GTP binding / nucleolus / perinuclear region of cytoplasm / Golgi apparatus / ATP hydrolysis activity / DNA binding / RNA binding / zinc ion binding / ATP binding / nucleus / membrane / plasma membrane / cytosol / cytoplasm Similarity search - Function | ||||||||||||
| Biological species | ![]() | ||||||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 2.9 Å | ||||||||||||
Authors | Nguyen TT / Hashem Y / Rocha REO / Boissier F / Qian SB / Jia L / Uematsu S / Gu Y / Shi S | ||||||||||||
| Funding support | United States, European Union, 3 items
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Citation | Journal: Nat Biotechnol / Year: 2025Title: Programmable initiation of mRNA translation by trans-RNA. Authors: Longfei Jia / Tan-Trung Nguyen / Saori Uematsu / Yifei Gu / Shengcho Shi / Yaser Hashem / Shu-Bing Qian / ![]() Abstract: Several approaches exist to silence genes, but few tools are available to activate individual mRNAs for translation inside cells. Guiding ribosomes to specific start codons without altering the ...Several approaches exist to silence genes, but few tools are available to activate individual mRNAs for translation inside cells. Guiding ribosomes to specific start codons without altering the original sequence remains a formidable task. Here we design capped trans-RNAs capable of directing ribosomes to specific initiation sites on individual mRNAs when the trans-cap is positioned near the target start codon. Structural and biochemical data suggest that the capped trans-RNA facilitates ribosome loading and scanning on the target mRNA through a synergistic mechanism involving alternative cap recognition. The trans-RNA also acts independently of the cap on the target mRNA, enabling translation of circular RNAs lacking internal ribosome entry sites. We apply trans-RNAs in vivo to achieve programmable alternative translation of endogenous genes in mouse liver. Finally, we provide the evidence for the existence of natural transcripts that, similarly to exogenous trans-RNAs, activate translation of endogenous mRNAs. | ||||||||||||
| History |
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Structure visualization
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Downloads & links
-EMDB archive
| Map data | emd_51903.map.gz | 450.4 MB | EMDB map data format | |
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| Header (meta data) | emd-51903-v30.xml emd-51903.xml | 70.7 KB 70.7 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_51903_fsc.xml | 16.5 KB | Display | FSC data file |
| Images | emd_51903.png | 71.5 KB | ||
| Filedesc metadata | emd-51903.cif.gz | 15.3 KB | ||
| Others | emd_51903_additional_1.map.gz emd_51903_half_map_1.map.gz emd_51903_half_map_2.map.gz | 397.8 MB 442.5 MB 442.5 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-51903 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-51903 | HTTPS FTP |
-Validation report
| Summary document | emd_51903_validation.pdf.gz | 1.1 MB | Display | EMDB validaton report |
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| Full document | emd_51903_full_validation.pdf.gz | 1.1 MB | Display | |
| Data in XML | emd_51903_validation.xml.gz | 25.7 KB | Display | |
| Data in CIF | emd_51903_validation.cif.gz | 33.9 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-51903 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-51903 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 9h6yMC ![]() 9h74C M: atomic model generated by this map C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_51903.map.gz / Format: CCP4 / Size: 476.8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | main map of the mammalian late-stage 48S initiation complex | ||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 0.93 Å | ||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Additional map: EMhanced map of the mammalian late-stage 48S initiation complex
| File | emd_51903_additional_1.map | ||||||||||||
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| Annotation | EMhanced map of the mammalian late-stage 48S initiation complex | ||||||||||||
| Projections & Slices |
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| Density Histograms |
-Half map: half map B of the mammalian late-stage 48S initiation complex
| File | emd_51903_half_map_1.map | ||||||||||||
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| Annotation | half map B of the mammalian late-stage 48S initiation complex | ||||||||||||
| Projections & Slices |
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| Density Histograms |
-Half map: half map A of the mammalian late-stage 48S initiation complex
| File | emd_51903_half_map_2.map | ||||||||||||
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| Annotation | half map A of the mammalian late-stage 48S initiation complex | ||||||||||||
| Projections & Slices |
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| Density Histograms |
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Sample components
+Entire : Late-stage 48S Initiation Complex (LS48S IC) guided by the trans-...
+Supramolecule #1: Late-stage 48S Initiation Complex (LS48S IC) guided by the trans-...
+Macromolecule #1: Initiator methionylated tRNA
+Macromolecule #2: 18S ribosomal RNA
+Macromolecule #3: Linear mRNA
+Macromolecule #4: Eukaryotic translation initiation factor 2 subunit 1
+Macromolecule #5: Eukaryotic translation initiation factor 2 subunit 3
+Macromolecule #6: Small ribosomal subunit protein uS2
+Macromolecule #7: Small ribosomal subunit protein eS1
+Macromolecule #8: Small ribosomal subunit protein uS5
+Macromolecule #9: Small ribosomal subunit protein uS3
+Macromolecule #10: Small ribosomal subunit protein eS4
+Macromolecule #11: Small ribosomal subunit protein uS7
+Macromolecule #12: Small ribosomal subunit protein eS6
+Macromolecule #13: Small ribosomal subunit protein eS7
+Macromolecule #14: Small ribosomal subunit protein eS8
+Macromolecule #15: Small ribosomal subunit protein uS4
+Macromolecule #16: Small ribosomal subunit protein eS10
+Macromolecule #17: Small ribosomal subunit protein uS17
+Macromolecule #18: Small ribosomal subunit protein eS12
+Macromolecule #19: Small ribosomal subunit protein uS15
+Macromolecule #20: Small ribosomal subunit protein uS11
+Macromolecule #21: Small ribosomal subunit protein uS19
+Macromolecule #22: Small ribosomal subunit protein uS9
+Macromolecule #23: Small ribosomal subunit protein eS17
+Macromolecule #24: Small ribosomal subunit protein uS13
+Macromolecule #25: Small ribosomal subunit protein eS19
+Macromolecule #26: Small ribosomal subunit protein uS10
+Macromolecule #27: Small ribosomal subunit protein eS21
+Macromolecule #28: Small ribosomal subunit protein uS8
+Macromolecule #29: Small ribosomal subunit protein uS12
+Macromolecule #30: 40S ribosomal protein S24
+Macromolecule #31: Small ribosomal subunit protein eS26
+Macromolecule #32: Small ribosomal subunit protein eS27
+Macromolecule #33: Small ribosomal subunit protein eS28
+Macromolecule #34: Small ribosomal subunit protein uS14
+Macromolecule #35: Ubiquitin-ribosomal protein eS31 fusion protein
+Macromolecule #36: Small ribosomal subunit protein RACK1
+Macromolecule #37: Ubiquitin-like FUBI-ribosomal protein eS30 fusion protein
+Macromolecule #38: Eukaryotic translation initiation factor 4C
+Macromolecule #39: ATP binding cassette subfamily E member 1
+Macromolecule #40: Small ribosomal subunit protein eS32
+Macromolecule #41: Small ribosomal subunit protein eS25
+Macromolecule #42: Eukaryotic translation initiation factor 5
+Macromolecule #43: IRON/SULFUR CLUSTER
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Concentration | 0.4 mg/mL | ||||||||||||||||||
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| Buffer | pH: 7.4 Component:
Details: 10 mM HEPES-KOH pH 7.4, 5 mM Mg(OAc)2, 50 mM KOAc, 10 mM NH4Cl, 2 mM DTT and Roche cOmplete protease inhibitor cocktail | ||||||||||||||||||
| Grid | Model: Quantifoil R2/2 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: HOLEY / Support film - Film thickness: 2 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 20 sec. / Details: glow-discharged at 2.5 mA for 20 s | ||||||||||||||||||
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV Details: The sample was incubated on the grid for 30 s and then blotted with filter paper from both sides for 1.5 s with the blot force 5. | ||||||||||||||||||
| Details | late-stage 48S initiation translation complex was dissolved in 20 microlitre buffer containing 10 mM HEPES-KOH pH 7.4, 5 mM Mg(OAc)2, 50 mM KOAc, 10 mM NH4Cl, 2 mM DTT and Roche cOmplete protease inhibitor cocktail |
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Electron microscopy
| Microscope | FEI TALOS ARCTICA |
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| Temperature | Min: 90.0 K / Max: 95.0 K |
| Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Digitization - Dimensions - Width: 4096 pixel / Digitization - Dimensions - Height: 4096 pixel / Digitization - Frames/image: 1-40 / Number grids imaged: 1 / Number real images: 9524 / Average exposure time: 6.0 sec. / Average electron dose: 50.0 e/Å2 |
| Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | C2 aperture diameter: 30.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.5 µm / Nominal defocus min: 0.5 µm / Nominal magnification: 59000 |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Talos Arctica / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
| Initial model |
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| Details | The atomic model of LS48S IC complex (PDB: 8P09) was initially rigid body fitted in density using ChimeraX and further model building was done for mRNA, eIF5 N-terminal domain, eIF2gamma, eIF3d and eIF3 octamer in ChimeraX. Densities for eIF5 N-terminal domain, eIF2gamma, eIF3d and eIF3 octamer were observed in the map and therefore were included in the model. The 3D structure of the eIF5-N terminal domain was generated by Alphafold2 (PDB: G1TGW1). It was then accommodated into the model by aligning structurally to its counterparts in the yeast atomic model of the py48S-eIF5N pre-initiation complex (PDB: 6FYX). IF3d and eIF3 octamer were taken from the atomic model of LS48S IC complex with eIF3 proteins (PDB: 6YAM) and placed into density by rigid-body fitting. The atomic models with and without eIF3 proteins were inspected visually and performed stereochemical, torsional, angle refinement and optimization by using ISOLDE v1.6. Figures featuring cryo-EM densities and atomic models were visualized by using ChimeraX. | |||||||||||||||
| Refinement | Protocol: RIGID BODY FIT | |||||||||||||||
| Output model | ![]() PDB-9h6y: |
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Keywords
Authors
United States, European Union, 3 items
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FIELD EMISSION GUN





