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Yorodumi- PDB-9h74: Late-stage 48S Initiation Complex with eIF3 (LS48S-eIF3 IC) guide... -
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Open data
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Basic information
| Entry | Database: PDB / ID: 9h74 | ||||||||||||
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| Title | Late-stage 48S Initiation Complex with eIF3 (LS48S-eIF3 IC) guided by the trans-RNA | ||||||||||||
Components |
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Keywords | TRANSLATION / 48S late-stage initiation complex / Programmable mRNA translation initiation / trans-RNA / activation of mRNA 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 / viral translational termination-reinitiation / L13a-mediated translational silencing of Ceruloplasmin expression / SRP-dependent cotranslational protein targeting to membrane ...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 / viral translational termination-reinitiation / 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 3 complex, eIF3e / cap-dependent translational initiation / eukaryotic translation initiation factor 3 complex, eIF3m / IRES-dependent viral translational initiation / translation reinitiation / eukaryotic translation initiation factor 2 complex / formation of cytoplasmic translation initiation complex / eukaryotic translation initiation factor 3 complex / multi-eIF complex / translation factor activity, RNA binding / eukaryotic 43S preinitiation complex / formation of translation preinitiation complex / mRNA cap binding / eukaryotic 48S preinitiation complex / cellular response to chemical stress / protein-synthesizing GTPase / metal-dependent deubiquitinase activity / nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / regulation of translational initiation / GDP-dissociation inhibitor activity / Formation of the ternary complex, and subsequently, the 43S complex / laminin receptor activity / Ribosomal scanning and start codon recognition / Translation initiation complex formation / Formation of a pool of free 40S subunits / 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 / GTP hydrolysis and joining of the 60S ribosomal subunit / L13a-mediated translational silencing of Ceruloplasmin expression / phagocytic cup / 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) / laminin binding / rough endoplasmic reticulum / translation regulator activity / ribosomal small subunit export from nucleus / translation initiation factor binding / gastrulation / translation initiation factor activity / MDM2/MDM4 family protein binding / 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 translation / ribosome assembly / 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 / translational initiation / small-subunit processome / PML body / spindle / fibrillar center / cytoplasmic stress granule / rRNA processing / positive regulation of canonical Wnt signaling pathway / rhythmic process / 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 / cysteine-type deubiquitinase activity / mitochondrial inner membrane / rRNA binding / postsynaptic density / structural constituent of ribosome / ribosome / translation / ribonucleoprotein complex / cell division / DNA repair / mRNA binding / GTPase activity / apoptotic process / dendrite / synapse Similarity search - Function | ||||||||||||
| Biological species | ![]() | ||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.9 Å | ||||||||||||
Authors | Nguyen, T.T. / Hashem, Y. / Rocha, R.E.O. / Boissier, F. / Qian, S.B. / Jia, L. / Uematsu, S. / Gu, Y. / Shi, S. | ||||||||||||
| Funding support | United States, European Union, 3items
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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. | ||||||||||||
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Structure visualization
| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 9h74.cif.gz | 2.7 MB | Display | PDBx/mmCIF format |
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| PDB format | pdb9h74.ent.gz | Display | PDB format | |
| PDBx/mmJSON format | 9h74.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/h7/9h74 ftp://data.pdbj.org/pub/pdb/validation_reports/h7/9h74 | HTTPS FTP |
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-Related structure data
| Related structure data | ![]() 51911MC ![]() 9h6yC M: map data used to model this data C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
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Assembly
| Deposited unit | ![]()
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Components
-RNA chain , 3 types, 3 molecules 123
| #1: RNA chain | Mass: 24721.828 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
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| #2: RNA chain | Mass: 601015.688 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
| #3: RNA chain | Mass: 51665.809 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
-Eukaryotic translation initiation factor ... , 12 types, 12 molecules Ajmoqrstuvwy
| #4: Protein | Mass: 44692.543 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
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| #38: Protein | Mass: 16476.395 Da / Num. of mol.: 1 / Source method: isolated from a natural source Details: Uniprot: G1SYS4, Eukaryotic translation initiation factor 4C. Many last papers called this protein as Eukaryotic translation initiation factor 1A Source: (natural) ![]() |
| #41: Protein | Mass: 64959.684 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: G1SQV5 (162-526) / Source: (natural) ![]() |
| #43: Protein | Mass: 49131.520 Da / Num. of mol.: 1 / Source method: isolated from a natural source Details: G1TGW1 (2019-12-11 v2) (430 aa) and A0A5F9C5D3 (2019-12-11 v1) (398 aa). Different the C-ter of two protein isoform from the position 354 Source: (natural) ![]() |
| #44: Protein | Mass: 37846.730 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: G1SLC2 (364 aa) (86-357) A0A5F9CZR4 (366 aa) / Source: (natural) ![]() |
| #45: Protein | Mass: 39955.324 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: A0A1U7U441 / Source: (natural) ![]() |
| #46: Protein | Mass: 25129.709 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: G1T3L2 / Source: (natural) ![]() |
| #47: Protein | Mass: 71001.477 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: G1SED9 (607 aa) (181-552) A0A5F9DP35 (466 aa) / Source: (natural) ![]() |
| #48: Protein | Mass: 42555.832 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: G1SLW8 / Source: (natural) ![]() |
| #49: Protein | Mass: 105676.172 Da / Num. of mol.: 1 / Source method: isolated from a natural source Details: G1U971 (913 aa) A0A5F9CMJ3 (892 aa). The difference is in N-terminal of two version of protein Eukaryotic translation initiation factor 3 subunit C (A0A5F9CMJ3 - 892 aa - position 1-10 vs ...Details: G1U971 (913 aa) A0A5F9CMJ3 (892 aa). The difference is in N-terminal of two version of protein Eukaryotic translation initiation factor 3 subunit C (A0A5F9CMJ3 - 892 aa - position 1-10 vs G1U971 - 913 aa - position position 1-31) Source: (natural) ![]() |
| #50: Protein | Mass: 54199.863 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: G1SUC8 (4-422) / Source: (natural) ![]() |
| #51: Protein | Mass: 164902.656 Da / Num. of mol.: 1 / Source method: isolated from a natural source Details: G1SMZ5 (1362 aa) A0A5F9C991 (1350 aa) A0A5F9CMG0 (1343 aa) A0A5F9C3S1 (1351 aa) The difference is in N-terminal of two version of protein Eukaryotic translation initiation factor 3 subunit A Source: (natural) ![]() |
-Protein , 2 types, 2 molecules Bk
| #5: Protein | Mass: 52499.992 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Uniprot ID: A0A5F9C3W2 / Source: (natural) ![]() References: UniProt: A0A5F9C3W2, protein-synthesizing GTPase |
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| #39: Protein | Mass: 67419.258 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: G1SG72 / Source: (natural) ![]() |
+40S ribosomal protein ... , 23 types, 23 molecules CDGJLNOPQRTUVWXZabcdein
-Small ribosomal subunit protein ... , 5 types, 5 molecules EIKMg
| #8: Protein | Mass: 29099.955 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: A0AAA9WW19 / Source: (natural) ![]() |
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| #12: Protein | Mass: 28755.895 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: G1TM55 / Source: (natural) ![]() |
| #14: Protein | Mass: 24163.244 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: G1TJW1 / Source: (natural) ![]() |
| #16: Protein | Mass: 17156.865 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: G1U0Y5 / Source: (natural) ![]() |
| #36: Protein | Mass: 35115.652 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: G1SJB4 / Source: (natural) ![]() |
-Ribosomal protein ... , 5 types, 5 molecules FHSYf
| #9: Protein | Mass: 26715.344 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: G1TNM3 / Source: (natural) ![]() |
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| #11: Protein | Mass: 22913.453 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: G1TFM5 / Source: (natural) ![]() |
| #22: Protein | Mass: 16477.377 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: G1SGX4 / Source: (natural) ![]() |
| #28: Protein | Mass: 14865.555 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: G1TG89 / Source: (natural) ![]() |
| #35: Protein | Mass: 18004.041 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: G1SK22 / Source: (natural) ![]() |
-Protein/peptide / Non-polymers , 2 types, 3 molecules l

| #40: Protein/peptide | Mass: 3473.451 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: A0A087WNH4 / Source: (natural) ![]() |
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| #52: Chemical |
-Details
| Has ligand of interest | Y |
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| Has protein modification | N |
-Experimental details
-Experiment
| Experiment | Method: ELECTRON MICROSCOPY |
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| EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
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Sample preparation
| Component | Name: Late-stage 48S Initiation Complex with eIF3 (LS48S-eIF3 IC) guided by the trans-RNA of 15 nt length, purification method of biotinylated DNA-oligo pull-down Type: RIBOSOME / Entity ID: #1-#51 / Source: NATURAL | ||||||||||||||||||||||||||||||
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| Molecular weight | Value: 1.5 MDa / Experimental value: NO | ||||||||||||||||||||||||||||||
| Source (natural) | Organism: ![]() | ||||||||||||||||||||||||||||||
| Buffer solution | pH: 7.4 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 | ||||||||||||||||||||||||||||||
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| Specimen | Conc.: 0.4 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES Details: late-stage 48S initiation translation complex with eIF3 (LS48S-eIF3 IC) 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 ...Details: late-stage 48S initiation translation complex with eIF3 (LS48S-eIF3 IC) 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 | ||||||||||||||||||||||||||||||
| Specimen support | Details: glow-discharged at 2.5 mA for 20 s / Grid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R2/2 | ||||||||||||||||||||||||||||||
| Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277 K 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 |
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Electron microscopy imaging
| Experimental equipment | ![]() Model: Talos Arctica / Image courtesy: FEI Company |
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| Microscopy | Model: FEI TALOS ARCTICA |
| Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 200 kV / Illumination mode: FLOOD BEAM |
| Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 59000 X / Nominal defocus max: 2500 nm / Nominal defocus min: 500 nm / Cs: 2.7 mm / C2 aperture diameter: 30 µm / Alignment procedure: COMA FREE |
| Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Temperature (max): 95 K / Temperature (min): 90 K |
| Image recording | Average exposure time: 6 sec. / Electron dose: 50 e/Å2 / Detector mode: COUNTING / Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Num. of grids imaged: 1 / Num. of real images: 9524 |
| Image scans | Sampling size: 6.35 µm / Width: 4096 / Height: 4096 / Movie frames/image: 40 / Used frames/image: 1-40 |
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Processing
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| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | |||||||||||||||||||||||||||||||||||||||||||||
| 3D reconstruction | Resolution: 2.9 Å / Resolution method: OTHER / Num. of particles: 1000 / Symmetry type: POINT | |||||||||||||||||||||||||||||||||||||||||||||
| Atomic model building | Protocol: RIGID BODY FIT 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, ...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. | |||||||||||||||||||||||||||||||||||||||||||||
| Atomic model building | 3D fitting-ID: 1
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