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Yorodumi- EMDB-25531: Structure of the HCV IRES binding to the 40S ribosomal subunit, c... -
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Open data
- Basic information
Basic information
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| Title | Structure of the HCV IRES binding to the 40S ribosomal subunit, closed conformation. Structure 5(delta dII) | |||||||||||||||
|  Map data | Post processed map | |||||||||||||||
|  Sample | 
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|  Keywords | HCV / IRES / 40S / RIBOSOME | |||||||||||||||
| Function / homology |  Function and homology information ribosomal subunit / laminin receptor activity / ubiquitin ligase inhibitor activity / positive regulation of signal transduction by p53 class mediator / 90S preribosome / phagocytic cup / ribosomal small subunit export from nucleus / laminin binding / rough endoplasmic reticulum / translation regulator activity ...ribosomal subunit / laminin receptor activity / ubiquitin ligase inhibitor activity / positive regulation of signal transduction by p53 class mediator / 90S preribosome / phagocytic cup / ribosomal small subunit export from nucleus / laminin binding / rough endoplasmic reticulum / translation regulator activity / gastrulation / MDM2/MDM4 family protein binding / cytosolic ribosome / class I DNA-(apurinic or apyrimidinic site) endonuclease activity / DNA-(apurinic or apyrimidinic site) lyase / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / positive regulation of apoptotic signaling pathway / maturation of SSU-rRNA / small-subunit processome / spindle / 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 / mitochondrial inner membrane / postsynaptic density / rRNA binding / structural constituent of ribosome / ribosome / translation / ribonucleoprotein complex / cell division / DNA repair / apoptotic process / dendrite / synapse / centrosome / nucleolus / perinuclear region of cytoplasm / Golgi apparatus / DNA binding / RNA binding / zinc ion binding / nucleus / membrane / plasma membrane / cytoplasm Similarity search - Function | |||||||||||||||
| Biological species |   Oryctolagus cuniculus (rabbit) /  hepatitis C virus genotype 1a | |||||||||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 4.2 Å | |||||||||||||||
|  Authors | Brown ZP / Abaeva IS / De S / Hellen CUT / Pestova TV / Frank J | |||||||||||||||
| Funding support |  United States, 4 items 
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|  Citation |  Journal: EMBO J / Year: 2022 Title: Molecular architecture of 40S translation initiation complexes on the hepatitis C virus IRES. Authors: Zuben P Brown / Irina S Abaeva / Swastik De / Christopher U T Hellen / Tatyana V Pestova / Joachim Frank /  Abstract: Hepatitis C virus mRNA contains an internal ribosome entry site (IRES) that mediates end-independent translation initiation, requiring a subset of eukaryotic initiation factors (eIFs). Biochemical ...Hepatitis C virus mRNA contains an internal ribosome entry site (IRES) that mediates end-independent translation initiation, requiring a subset of eukaryotic initiation factors (eIFs). Biochemical studies revealed that direct binding of the IRES to the 40S ribosomal subunit places the initiation codon into the P site, where it base pairs with eIF2-bound Met-tRNAiMet forming a 48S initiation complex. Subsequently, eIF5 and eIF5B mediate subunit joining, yielding an elongation-competent 80S ribosome. Initiation can also proceed without eIF2, in which case Met-tRNAiMet is recruited directly by eIF5B. However, the structures of initiation complexes assembled on the HCV IRES, the transitions between different states, and the accompanying conformational changes have remained unknown. To fill these gaps, we now obtained cryo-EM structures of IRES initiation complexes, at resolutions up to 3.5 Å, that cover all major stages from the initial ribosomal association, through eIF2-containing 48S initiation complexes, to eIF5B-containing complexes immediately prior to subunit joining. These structures provide insights into the dynamic network of 40S/IRES contacts, highlight the role of IRES domain II, and reveal conformational changes that occur during the transition from eIF2- to eIF5B-containing 48S complexes and prepare them for subunit joining. | |||||||||||||||
| History | 
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- Structure visualization
Structure visualization
| Supplemental images | 
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- Downloads & links
Downloads & links
-EMDB archive
| Map data |  emd_25531.map.gz | 227.9 MB |  EMDB map data format | |
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| Header (meta data) |  emd-25531-v30.xml  emd-25531.xml | 60.2 KB 60.2 KB | Display Display |  EMDB header | 
| FSC (resolution estimation) |  emd_25531_fsc.xml | 14.3 KB | Display |  FSC data file | 
| Images |  emd_25531.png | 35.9 KB | ||
| Masks |  emd_25531_msk_1.map | 244.1 MB |  Mask map | |
| Filedesc metadata |  emd-25531.cif.gz | 12.5 KB | ||
| Others |  emd_25531_additional_1.map.gz  emd_25531_additional_2.map.gz  emd_25531_additional_3.map.gz  emd_25531_half_map_1.map.gz  emd_25531_half_map_2.map.gz | 144.8 MB 122.6 MB 193.6 MB 194.1 MB 193.7 MB | ||
| Archive directory |  http://ftp.pdbj.org/pub/emdb/structures/EMD-25531  ftp://ftp.pdbj.org/pub/emdb/structures/EMD-25531 | HTTPS FTP | 
-Validation report
| Summary document |  emd_25531_validation.pdf.gz | 1.2 MB | Display |  EMDB validaton report | 
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| Full document |  emd_25531_full_validation.pdf.gz | 1.2 MB | Display | |
| Data in XML |  emd_25531_validation.xml.gz | 20.8 KB | Display | |
| Data in CIF |  emd_25531_validation.cif.gz | 27 KB | Display | |
| Arichive directory |  https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-25531  ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-25531 | HTTPS FTP | 
-Related structure data
| Related structure data |  7sykMC  7syiC  7syjC  7sylC  7syoC  7sypC  7syqC  7syrC  7sysC  7sytC  7syuC  7syvC  7sywC  7syxC 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 | 
- Links
Links
| EMDB pages |  EMDB (EBI/PDBe) /  EMDataResource | 
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| Related items in Molecule of the Month | 
- Map
Map
| File |  Download / File: emd_25531.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | Post processed map | ||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
 
 Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 0.95 Å | ||||||||||||||||||||||||||||||||||||
| Density | 
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML: 
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-Supplemental data
-Mask #1
| File |  emd_25531_msk_1.map | ||||||||||||
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| Projections & Slices | 
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| Density Histograms | 
-Additional map: Local resolution map filtered at local resolution
| File | emd_25531_additional_1.map | ||||||||||||
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| Annotation | Local resolution map filtered at local resolution | ||||||||||||
| Projections & Slices | 
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| Density Histograms | 
-Additional map: Local resolution values
| File | emd_25531_additional_2.map | ||||||||||||
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| Annotation | Local resolution values | ||||||||||||
| Projections & Slices | 
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| Density Histograms | 
-Additional map: Unsharpened map
| File | emd_25531_additional_3.map | ||||||||||||
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| Annotation | Unsharpened map | ||||||||||||
| Projections & Slices | 
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| Density Histograms | 
-Half map: Half map
| File | emd_25531_half_map_1.map | ||||||||||||
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| Annotation | Half map | ||||||||||||
| Projections & Slices | 
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| Density Histograms | 
-Half map: Half map
| File | emd_25531_half_map_2.map | ||||||||||||
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| Annotation | Half map | ||||||||||||
| Projections & Slices | 
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| Density Histograms | 
- Sample components
Sample components
+Entire : 40S ribosomal small subunit with HCV IRES
+Supramolecule #1: 40S ribosomal small subunit with HCV IRES
+Macromolecule #1: 18S rRNA
+Macromolecule #36: HCV IRES
+Macromolecule #2: 40S ribosomal protein SA
+Macromolecule #3: eS1
+Macromolecule #4: uS5 (S2)
+Macromolecule #5: uS3
+Macromolecule #6: 40S ribosomal protein S4
+Macromolecule #7: uS7
+Macromolecule #8: eS6
+Macromolecule #9: 40S ribosomal protein S7
+Macromolecule #10: eS8
+Macromolecule #11: uS4
+Macromolecule #12: eS10
+Macromolecule #13: uS17
+Macromolecule #14: eS12
+Macromolecule #15: uS15
+Macromolecule #16: uS11
+Macromolecule #17: uS19
+Macromolecule #18: uS9
+Macromolecule #19: eS17
+Macromolecule #20: uS13
+Macromolecule #21: eS19
+Macromolecule #22: uS10
+Macromolecule #23: 40S ribosomal protein S21
+Macromolecule #24: uS8
+Macromolecule #25: uS12
+Macromolecule #26: 40S ribosomal protein S24
+Macromolecule #27: eS25 (S25)
+Macromolecule #28: eS26 (S26)
+Macromolecule #29: eS27
+Macromolecule #30: eS28
+Macromolecule #31: eS29
+Macromolecule #32: eS30
+Macromolecule #33: 40S ribosomal protein S27a
+Macromolecule #34: RACK1
+Macromolecule #35: 60s ribosomal protein l41
+Macromolecule #37: ZINC ION
-Experimental details
-Structure determination
| Method | cryo EM | 
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|  Processing | single particle reconstruction | 
| Aggregation state | particle | 
- Sample preparation
Sample preparation
| Concentration | 0.000075 mg/mL | 
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| Buffer | pH: 7.5 | 
| Grid | Model: Quantifoil R0.6/1 / Material: GOLD / Mesh: 300 / Support film - Material: GOLD / Support film - topology: HOLEY / Support film - Film thickness: 50 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 25 sec. / Pretreatment - Atmosphere: OTHER / Details: H2/O2 mixture for 25 seconds at 25W power | 
| Vitrification | Cryogen name: ETHANE-PROPANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV / Details: 4 second blot time, force 3. | 
- Electron microscopy
Electron microscopy
| Microscope | FEI TECNAI F30 | 
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| Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average exposure time: 4.0 sec. / Average electron dose: 70.9 e/Å2 | 
| Electron beam | Acceleration voltage: 300 kV / Electron source:  FIELD EMISSION GUN | 
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.26 mm / Nominal defocus max: 2.0 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 52000 | 
| Sample stage | Cooling holder cryogen: NITROGEN | 
| Experimental equipment |  Model: Tecnai F30 / Image courtesy: FEI Company | 
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