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Yorodumi- EMDB-20256: Structure of a mammalian 80S ribosome in complex with the Israeli... -
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Open data
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Basic information
| Entry | Database: EMDB / ID: EMD-20256 | |||||||||
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| Title | Structure of a mammalian 80S ribosome in complex with the Israeli Acute Paralysis Virus IRES (Class 2) | |||||||||
Map data | mammalian 80S ribosome in complex with the Israeli Acute Paralysis Virus IRES (Class 2) | |||||||||
Sample |
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Keywords | Israeli Acute Paralysis Virus / Internal Ribosome Entry Site / IRES / Small Ribosomal Subunit / 40S / Large Ribosomal Subunit / 60S / 80S / ribosomes / RIBOSOME | |||||||||
| Function / homology | Function and homology informationribosomal subunit / ubiquitin ligase inhibitor activity / positive regulation of signal transduction by p53 class mediator / 90S preribosome / phagocytic cup / ribosomal small subunit export from nucleus / rough endoplasmic reticulum / translation regulator activity / gastrulation / MDM2/MDM4 family protein binding ...ribosomal subunit / ubiquitin ligase inhibitor activity / positive regulation of signal transduction by p53 class mediator / 90S preribosome / phagocytic cup / ribosomal small subunit export from nucleus / 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 LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / ribosomal large subunit biogenesis / 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 / antimicrobial humoral immune response mediated by antimicrobial peptide / rhythmic process / positive regulation of canonical Wnt signaling pathway / heparin binding / large ribosomal subunit / regulation of translation / ribosome binding / ribosomal small subunit biogenesis / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / small ribosomal subunit rRNA binding / ribosomal large subunit assembly / cytosolic small ribosomal subunit / large ribosomal subunit rRNA binding / perikaryon / defense response to Gram-negative bacterium / killing of cells of another organism / cytosolic large ribosomal subunit / cytoplasmic translation / cell differentiation / tRNA binding / mitochondrial inner membrane / postsynaptic density / rRNA binding / structural constituent of ribosome / ribosome / translation / ribonucleoprotein complex / cell division / DNA repair / mRNA binding / apoptotic process / dendrite / synapse / centrosome / nucleolus / perinuclear region of cytoplasm / endoplasmic reticulum / Golgi apparatus / DNA binding / RNA binding / zinc ion binding / nucleus / cytoplasm / cytosol Similarity search - Function | |||||||||
| Biological species | ![]() Israeli acute paralysis virus | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.1 Å | |||||||||
Authors | Acosta-Reyes FJ / Neupane R | |||||||||
| Funding support | United States, 1 items
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Citation | Journal: EMBO J / Year: 2019Title: The Israeli acute paralysis virus IRES captures host ribosomes by mimicking a ribosomal state with hybrid tRNAs. Authors: Francisco Acosta-Reyes / Ritam Neupane / Joachim Frank / Israel S Fernández / ![]() Abstract: Colony collapse disorder (CCD) is a multi-faceted syndrome decimating bee populations worldwide, and a group of viruses of the widely distributed Dicistroviridae family have been identified as a ...Colony collapse disorder (CCD) is a multi-faceted syndrome decimating bee populations worldwide, and a group of viruses of the widely distributed Dicistroviridae family have been identified as a causing agent of CCD. This family of viruses employs non-coding RNA sequences, called internal ribosomal entry sites (IRESs), to precisely exploit the host machinery for viral protein production. Using single-particle cryo-electron microscopy (cryo-EM), we have characterized how the IRES of Israeli acute paralysis virus (IAPV) intergenic region captures and redirects translating ribosomes toward viral RNA messages. We reconstituted two in vitro reactions targeting a pre-translocation and a post-translocation state of the IAPV-IRES in the ribosome, allowing us to identify six structures using image processing classification methods. From these, we reconstructed the trajectory of IAPV-IRES from the early small subunit recruitment to the final post-translocated state in the ribosome. An early commitment of IRES/ribosome complexes for global pre-translocation mimicry explains the high efficiency observed for this IRES. Efforts directed toward fighting CCD by targeting the IAPV-IRES using RNA-interference technology are underway, and the structural framework presented here may assist in further refining these approaches. | |||||||||
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Structure visualization
| Movie |
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| Structure viewer | EM map: SurfView Molmil Jmol/JSmol |
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_20256.map.gz | 27.1 MB | EMDB map data format | |
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| Header (meta data) | emd-20256-v30.xml emd-20256.xml | 103.7 KB 103.7 KB | Display Display | EMDB header |
| Images | emd_20256.png | 216.3 KB | ||
| Masks | emd_20256_msk_1.map | 178 MB | Mask map | |
| Filedesc metadata | emd-20256.cif.gz | 20.3 KB | ||
| Others | emd_20256_additional.map.gz emd_20256_half_map_1.map.gz emd_20256_half_map_2.map.gz | 140.7 MB 141.1 MB 141.2 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-20256 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-20256 | HTTPS FTP |
-Validation report
| Summary document | emd_20256_validation.pdf.gz | 1 MB | Display | EMDB validaton report |
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| Full document | emd_20256_full_validation.pdf.gz | 1 MB | Display | |
| Data in XML | emd_20256_validation.xml.gz | 14.9 KB | Display | |
| Data in CIF | emd_20256_validation.cif.gz | 17.8 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-20256 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-20256 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 6p5jMC ![]() 6p4gC ![]() 6p4hC ![]() 6p5iC ![]() 6p5kC ![]() 6p5nC C: citing same article ( M: atomic model generated by this map |
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| Similar structure data |
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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_20256.map.gz / Format: CCP4 / Size: 178 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Annotation | mammalian 80S ribosome in complex with the Israeli Acute Paralysis Virus IRES (Class 2) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.233 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
| File | emd_20256_msk_1.map | ||||||||||||
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| Projections & Slices |
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| Density Histograms |
-Additional map: mammalian 80S ribosome in complex with the Israeli...
| File | emd_20256_additional.map | ||||||||||||
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| Annotation | mammalian 80S ribosome in complex with the Israeli Acute Paralysis Virus IRES (Class 2) | ||||||||||||
| Projections & Slices |
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| Density Histograms |
-Half map: mammalian 80S ribosome in complex with the Israeli...
| File | emd_20256_half_map_1.map | ||||||||||||
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| Annotation | mammalian 80S ribosome in complex with the Israeli Acute Paralysis Virus IRES (Class 2) | ||||||||||||
| Projections & Slices |
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| Density Histograms |
-Half map: mammalian 80S ribosome in complex with the Israeli...
| File | emd_20256_half_map_2.map | ||||||||||||
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| Annotation | mammalian 80S ribosome in complex with the Israeli Acute Paralysis Virus IRES (Class 2) | ||||||||||||
| Projections & Slices |
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| Density Histograms |
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Sample components
+Entire : Structure of a mammalian 80S ribosome in complex with the Israeli...
+Supramolecule #1: Structure of a mammalian 80S ribosome in complex with the Israeli...
+Macromolecule #1: 18S rRNA
+Macromolecule #35: IAPV-IRES
+Macromolecule #36: 28S rRNA
+Macromolecule #37: 5S rRNA
+Macromolecule #38: 5.8S rRNA
+Macromolecule #2: uS2
+Macromolecule #3: eS1
+Macromolecule #4: uS5
+Macromolecule #5: uS3
+Macromolecule #6: eS4
+Macromolecule #7: uS7
+Macromolecule #8: eS6
+Macromolecule #9: eS7
+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: eS21
+Macromolecule #24: uS8
+Macromolecule #25: uS12
+Macromolecule #26: eS24
+Macromolecule #27: eS25
+Macromolecule #28: eS26
+Macromolecule #29: eS27
+Macromolecule #30: eS28
+Macromolecule #31: eS29
+Macromolecule #32: eS30
+Macromolecule #33: eS31
+Macromolecule #34: RACK1
+Macromolecule #39: uL2
+Macromolecule #40: uL3
+Macromolecule #41: uL4
+Macromolecule #42: uL18
+Macromolecule #43: eL6
+Macromolecule #44: uL30
+Macromolecule #45: eL8
+Macromolecule #46: uL6
+Macromolecule #47: uL16
+Macromolecule #48: uL11
+Macromolecule #49: eL13
+Macromolecule #50: L14e
+Macromolecule #51: eL15
+Macromolecule #52: uL13
+Macromolecule #53: uL22
+Macromolecule #54: eL18
+Macromolecule #55: eL19
+Macromolecule #56: eL20
+Macromolecule #57: eL21
+Macromolecule #58: eL22
+Macromolecule #59: uL14
+Macromolecule #60: eL24
+Macromolecule #61: eL23
+Macromolecule #62: uL24
+Macromolecule #63: eL27
+Macromolecule #64: uL15
+Macromolecule #65: eL29
+Macromolecule #66: eL30
+Macromolecule #67: eL31
+Macromolecule #68: eL32
+Macromolecule #69: eL33
+Macromolecule #70: eL34
+Macromolecule #71: eL35
+Macromolecule #72: eL36
+Macromolecule #73: eL37
+Macromolecule #74: eL38
+Macromolecule #75: eL39
+Macromolecule #76: eL40
+Macromolecule #77: eL41
+Macromolecule #78: eL42
+Macromolecule #79: eL43
+Macromolecule #80: eL28
+Macromolecule #81: uL1
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Buffer | pH: 7.5 Component:
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| Grid | Model: Quantifoil R2/2 / Material: COPPER / Support film - Material: CARBON / Support film - topology: HOLEY / Support film - Film thickness: 5 / Pretreatment - Type: PLASMA CLEANING / Pretreatment - Time: 25 sec. / Pretreatment - Atmosphere: OTHER / Pretreatment - Pressure: 9.33257 kPa Details: Plasma cleaning for both holey carbon and holey gold grids was done on a Gatan Solarus with Hydrogen (6.4 sccm gas flow) and Oxygen (27.5 sccm gas flow) and 10 W cleaning power | ||||||||||
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV Details: Blot force = 3s Wait time = 15s Drain time = 0s Blot time = 2.5 to 3 s. | ||||||||||
| Details | Ribosomal complexes for the pre-translocated state were assembled at 240-390 nM concentration and applied to plasma treated holey carbon. |
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Electron microscopy
| Microscope | FEI TECNAI F30 |
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| Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Digitization - Dimensions - Width: 3710 pixel / Digitization - Dimensions - Height: 3838 pixel / Digitization - Frames/image: 1-40 / Number real images: 11234 / Average exposure time: 8.0 sec. / Average electron dose: 42.09 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: OTHER / Imaging mode: BRIGHT FIELD / Cs: 2.26 mm / Nominal defocus max: 2.5 µm / Nominal defocus min: 0.8 µm / Nominal magnification: 31000 |
| Sample stage | Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Tecnai F30 / Image courtesy: FEI Company |
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About Yorodumi


Keywords
Israeli acute paralysis virus
Authors
United States, 1 items
Citation
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