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Yorodumi- PDB-4ujd: mammalian 80S HCV-IRES initiation complex with eIF5B PRE-like state -
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Open data
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Basic information
| Entry | Database: PDB / ID: 4ujd | ||||||||||||
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| Title | mammalian 80S HCV-IRES initiation complex with eIF5B PRE-like state | ||||||||||||
Components |
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Keywords | RIBOSOME / TRANSLATION INITIATION / HEPATITIS C VIRUS INTERNAL RIBOSOME ENTRY SITE / EUKARYOTIC INITIATION FACTOR 5B | ||||||||||||
| Function / homology | Function and homology informationprotein-synthesizing GTPase / translation initiation factor activity / GTPase activity / GTP binding / mitochondrion Similarity search - Function | ||||||||||||
| Biological species | ![]() HEPATITIS C VIRUS | ||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 8.9 Å | ||||||||||||
Authors | Yamamoto, H. / Unbehaun, A. / Loerke, J. / Behrmann, E. / Marianne, C. / Burger, J. / Mielke, T. / Spahn, C.M.T. | ||||||||||||
Citation | Journal: Nat Struct Mol Biol / Year: 2014Title: Structure of the mammalian 80S initiation complex with initiation factor 5B on HCV-IRES RNA. Authors: Hiroshi Yamamoto / Anett Unbehaun / Justus Loerke / Elmar Behrmann / Marianne Collier / Jörg Bürger / Thorsten Mielke / Christian M T Spahn / ![]() Abstract: The universally conserved eukaryotic initiation factor (eIF) 5B, a translational GTPase, is essential for canonical translation initiation. It is also required for initiation facilitated by the ...The universally conserved eukaryotic initiation factor (eIF) 5B, a translational GTPase, is essential for canonical translation initiation. It is also required for initiation facilitated by the internal ribosomal entry site (IRES) of hepatitis C virus (HCV) RNA. eIF5B promotes joining of 60S ribosomal subunits to 40S ribosomal subunits bound by initiator tRNA (Met-tRNAi(Met)). However, the exact molecular mechanism by which eIF5B acts has not been established. Here we present cryo-EM reconstructions of the mammalian 80S-HCV-IRES-Met-tRNAi(Met)-eIF5B-GMPPNP complex. We obtained two substates distinguished by the rotational state of the ribosomal subunits and the configuration of initiator tRNA in the peptidyl (P) site. Accordingly, a combination of conformational changes in the 80S ribosome and in initiator tRNA facilitates binding of the Met-tRNAi(Met) to the 60S P site and redefines the role of eIF5B as a tRNA-reorientation factor. | ||||||||||||
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Structure visualization
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| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 4ujd.cif.gz | 4.8 MB | Display | PDBx/mmCIF format |
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| PDB format | pdb4ujd.ent.gz | Display | PDB format | |
| PDBx/mmJSON format | 4ujd.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Summary document | 4ujd_validation.pdf.gz | 1.8 MB | Display | wwPDB validaton report |
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| Full document | 4ujd_full_validation.pdf.gz | 2.1 MB | Display | |
| Data in XML | 4ujd_validation.xml.gz | 341.4 KB | Display | |
| Data in CIF | 4ujd_validation.cif.gz | 609.4 KB | Display | |
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/uj/4ujd ftp://data.pdbj.org/pub/pdb/validation_reports/uj/4ujd | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 2682MC ![]() 2683C ![]() 4ujcC C: citing same article ( M: map data used to model this data |
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| Similar structure data |
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Links
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Assembly
| Deposited unit | ![]()
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Components
-RNA chain , 6 types, 6 molecules A2A3A4BABCC1
| #1: RNA chain | Mass: 1625917.625 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
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| #2: RNA chain | Mass: 62616.344 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
| #3: RNA chain | Mass: 38998.078 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
| #47: RNA chain | Mass: 24485.539 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
| #49: RNA chain | Mass: 162190.062 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) HEPATITIS C VIRUS |
| #50: RNA chain | Mass: 602776.875 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
+60S RIBOSOMAL PROTEIN ... , 42 types, 42 molecules AAABACADAEAFAGAHAIAJALAMANAOAPAQARASATAUAVAWAXAYAZAaAbAcAdAe...
-Protein , 2 types, 2 molecules AmBB
| #41: Protein | Mass: 14758.394 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
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| #48: Protein | Mass: 70788.000 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
+40S RIBOSOMAL PROTEIN ... , 33 types, 33 molecules CACBCCCDCECFCGCHCICJCKCLCMCNCOCPCQCRCSCTCUCVCWCXCYCZCaCbCcCd...
-Non-polymers , 2 types, 2 molecules 


| #84: Chemical | ChemComp-GNP / |
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| #85: Chemical | ChemComp-MG / |
-Details
| Has protein modification | Y |
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| Sequence details | CHAIN 2 1-5025 GB M11167 1-5025 CHAIN 3 1-181 GB CR749662 1-181 CHAIN 4 1-121 GB X51545 274-394 |
-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: HCV-IRES 80S INITIATION COMPLEX WITH EIF5B PRE- LIKE STATE Type: RIBOSOME |
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| Buffer solution | Name: 20MM TRIS-HCL, 5MM MGCL2, 100MM KCL, 0.2MM SPERMIDINE, 2MM DTT pH: 7.6 Details: 20MM TRIS-HCL, 5MM MGCL2, 100MM KCL, 0.2MM SPERMIDINE, 2MM DTT |
| Specimen | Conc.: 0.15 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
| Specimen support | Details: HOLEY CARBON |
| Vitrification | Instrument: FEI VITROBOT MARK II / Cryogen name: ETHANE Details: VITRIFICATION 1 -- CRYOGEN- ETHANE, HUMIDITY- 100, TEMPERATURE- 93, INSTRUMENT- FEI VITROBOT MARK II, METHOD- BLOT FOR 2-4 SECONDS BEFORE PLUNGING, |
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Electron microscopy imaging
| Experimental equipment | ![]() Model: Tecnai F30 / Image courtesy: FEI Company |
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| Microscopy | Model: FEI TECNAI F30 / Date: Dec 22, 2012 / Details: AUTOMATED DATA COLLECTION USING LEGINON |
| Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
| Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 115000 X / Calibrated magnification: 194805 X / Nominal defocus max: 4500 nm / Nominal defocus min: 2000 nm / Cs: 2 mm |
| Image recording | Electron dose: 20 e/Å2 / Film or detector model: TVIPS TEMCAM-F416 (4k x 4k) |
| Radiation wavelength | Relative weight: 1 |
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Processing
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| CTF correction | Details: CTFFIND3 | ||||||||||||
| Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||
| 3D reconstruction | Resolution: 8.9 Å / Num. of particles: 541570 / Nominal pixel size: 2.37 Å / Actual pixel size: 2.37 Å Details: SUBMISSION BASED ON EXPERIMENTAL DATA FROM EMDB EMD-2682. (DEPOSITION ID: 12562). Symmetry type: POINT | ||||||||||||
| Atomic model building | Protocol: RIGID BODY FIT / Space: REAL / Details: METHOD--RIGID BODY | ||||||||||||
| Atomic model building | PDB-ID: 4CXC![]() 4cxc Accession code: 4CXC / Source name: PDB / Type: experimental model | ||||||||||||
| Refinement | Highest resolution: 8.9 Å | ||||||||||||
| Refinement step | Cycle: LAST / Highest resolution: 8.9 Å
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About Yorodumi




HEPATITIS C VIRUS
Citation
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