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Yorodumi- EMDB-0058: Cryo-EM map of a partial yeast 48S preinitiation complex in its c... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-0058 | |||||||||
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Title | Cryo-EM map of a partial yeast 48S preinitiation complex in its closed conformation obtained by focused classification using masks for eIF3b-eIF3i. | |||||||||
Map data | For optimal visualization of eIF2, and of eIF3c, eIF3b and eIf3a c-term at the subunit interface, gauss-filter the map by 1.34 and display it at 0.035 | |||||||||
Sample |
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Function / homology | Function and homology information formation of translation initiation ternary complex / Recycling of eIF2:GDP / ABC-family proteins mediated transport / eukaryotic translation initiation factor 3 complex, eIF3e / methionyl-initiator methionine tRNA binding / eukaryotic translation initiation factor 3 complex, eIF3m / translation reinitiation / eukaryotic translation initiation factor 2 complex / incipient cellular bud site / multi-eIF complex ...formation of translation initiation ternary complex / Recycling of eIF2:GDP / ABC-family proteins mediated transport / eukaryotic translation initiation factor 3 complex, eIF3e / methionyl-initiator methionine tRNA binding / eukaryotic translation initiation factor 3 complex, eIF3m / translation reinitiation / eukaryotic translation initiation factor 2 complex / incipient cellular bud site / multi-eIF complex / eukaryotic translation initiation factor 3 complex / eukaryotic 43S preinitiation complex / cytoplasmic translational initiation / protein-synthesizing GTPase / formation of cytoplasmic translation initiation complex / formation of translation preinitiation complex / eukaryotic 48S preinitiation complex / positive regulation of translational fidelity / Formation of the ternary complex, and subsequently, the 43S complex / Translation initiation complex formation / Ribosomal scanning and start codon recognition / Formation of a pool of free 40S subunits / ribosomal small subunit binding / L13a-mediated translational silencing of Ceruloplasmin expression / translation regulator activity / regulation of translational fidelity / translation initiation factor binding / translational initiation / translation initiation factor activity / cytosolic ribosome / maintenance of translational fidelity / ribosomal small subunit assembly / rRNA processing / cytoplasmic stress granule / cytosolic small ribosomal subunit / ribosome binding / double-stranded RNA binding / small ribosomal subunit / cytosolic large ribosomal subunit / cytoplasmic translation / rRNA binding / ribosome / structural constituent of ribosome / translation / ribonucleoprotein complex / mRNA binding / GTPase activity / GTP binding / protein kinase binding / RNA binding / zinc ion binding / identical protein binding / metal ion binding / nucleus / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) / Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (yeast) / Yeast (fungus) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 5.75 Å | |||||||||
Authors | Llacer JL / Hussain T / Ramakrishnan V | |||||||||
Funding support | United Kingdom, 2 items
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Citation | Journal: Mol Cell / Year: 2015 Title: Conformational Differences between Open and Closed States of the Eukaryotic Translation Initiation Complex. Authors: Jose L Llácer / Tanweer Hussain / Laura Marler / Colin Echeverría Aitken / Anil Thakur / Jon R Lorsch / Alan G Hinnebusch / V Ramakrishnan / Abstract: Translation initiation in eukaryotes begins with the formation of a pre-initiation complex (PIC) containing the 40S ribosomal subunit, eIF1, eIF1A, eIF3, ternary complex (eIF2-GTP-Met-tRNAi), and ...Translation initiation in eukaryotes begins with the formation of a pre-initiation complex (PIC) containing the 40S ribosomal subunit, eIF1, eIF1A, eIF3, ternary complex (eIF2-GTP-Met-tRNAi), and eIF5. The PIC, in an open conformation, attaches to the 5' end of the mRNA and scans to locate the start codon, whereupon it closes to arrest scanning. We present single particle cryo-electron microscopy (cryo-EM) reconstructions of 48S PICs from yeast in these open and closed states, at 6.0 Å and 4.9 Å, respectively. These reconstructions show eIF2β as well as a configuration of eIF3 that appears to encircle the 40S, occupying part of the subunit interface. Comparison of the complexes reveals a large conformational change in the 40S head from an open mRNA latch conformation to a closed one that constricts the mRNA entry channel and narrows the P site to enclose tRNAi, thus elucidating key events in start codon recognition. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_0058.map.gz | 96.4 MB | EMDB map data format | |
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Header (meta data) | emd-0058-v30.xml emd-0058.xml | 77.3 KB 77.3 KB | Display Display | EMDB header |
Images | emd_0058.png | 75.3 KB | ||
Others | emd_0058_half_map_1.map.gz emd_0058_half_map_2.map.gz | 80.8 MB 80.8 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-0058 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-0058 | HTTPS FTP |
-Related structure data
Related structure data | 6gsnMC 0057C 6gsmC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_0058.map.gz / Format: CCP4 / Size: 103 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | For optimal visualization of eIF2, and of eIF3c, eIF3b and eIf3a c-term at the subunit interface, gauss-filter the map by 1.34 and display it at 0.035 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.34 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Half map: #1
File | emd_0058_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_0058_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Structure of a partial yeast 48S preinitiation complex in closed ...
+Supramolecule #1: Structure of a partial yeast 48S preinitiation complex in closed ...
+Supramolecule #3: tRNA
+Supramolecule #4: Initiation factors
+Supramolecule #5: mRNA
+Supramolecule #6: Initiation factors
+Supramolecule #2: Ribosome
+Macromolecule #1: 18S rRNA (1798-MER)
+Macromolecule #20: mRNA
+Macromolecule #28: tRNAi (75-MER)
+Macromolecule #2: KLLA0F09812p
+Macromolecule #3: KLLA0D08305p
+Macromolecule #4: KLLA0D10659p
+Macromolecule #5: KLLA0B08173p
+Macromolecule #6: 40S ribosomal protein S12
+Macromolecule #7: KLLA0F07843p
+Macromolecule #8: 40S ribosomal protein S16
+Macromolecule #9: KLLA0B01474p
+Macromolecule #10: KLLA0B01562p
+Macromolecule #11: KLLA0A07194p
+Macromolecule #12: KLLA0F25542p
+Macromolecule #13: KLLA0B06182p
+Macromolecule #14: 40S ribosomal protein S28
+Macromolecule #15: Ubiquitin-40S ribosomal protein S27a
+Macromolecule #16: KLLA0E12277p
+Macromolecule #17: 40S ribosomal protein S29
+Macromolecule #18: 40S ribosomal protein S30
+Macromolecule #19: 60S ribosomal protein L41-A
+Macromolecule #21: Eukaryotic translation initiation factor 1A
+Macromolecule #22: Eukaryotic translation initiation factor eIF-1
+Macromolecule #23: Eukaryotic translation initiation factor 3 subunit A,eIF3a
+Macromolecule #24: Eukaryotic translation initiation factor 3 subunit C
+Macromolecule #25: Eukaryotic translation initiation factor 2 subunit gamma
+Macromolecule #26: Eukaryotic translation initiation factor 2 subunit beta
+Macromolecule #27: Eukaryotic translation initiation factor 2 subunit alpha
+Macromolecule #29: 40S ribosomal protein S0
+Macromolecule #30: 40S ribosomal protein S1
+Macromolecule #31: 40S ribosomal protein S4
+Macromolecule #32: 40S ribosomal protein S6
+Macromolecule #33: 40S ribosomal protein S7
+Macromolecule #34: 40S ribosomal protein S8
+Macromolecule #35: KLLA0E23673p
+Macromolecule #36: KLLA0A10483p
+Macromolecule #37: KLLA0F18040p
+Macromolecule #38: 40S ribosomal protein S14
+Macromolecule #39: 40S ribosomal protein S21
+Macromolecule #40: 40S ribosomal protein S22
+Macromolecule #41: 40S ribosomal protein S24
+Macromolecule #42: RPS23
+Macromolecule #43: 40S ribosomal protein S26
+Macromolecule #44: 40S ribosomal protein S27
+Macromolecule #45: Eukaryotic translation initiation factor 3 subunit B
+Macromolecule #46: Eukaryotic translation initiation factor 3 subunit I
+Macromolecule #47: Eukaryotic translation initiation factor 3 subunit G
+Macromolecule #48: MAGNESIUM ION
+Macromolecule #49: ZINC ION
+Macromolecule #50: PHOSPHOMETHYLPHOSPHONIC ACID GUANYLATE ESTER
+Macromolecule #51: METHIONINE
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.18 mg/mL | ||||||||||||||
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Buffer | pH: 6.5 Component:
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Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK I |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal defocus max: 3.5 µm / Nominal defocus min: 1.5 µm / Nominal magnification: 59000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Temperature | Min: 90.0 K / Max: 100.0 K |
Image recording | Film or detector model: FEI FALCON II (4k x 4k) / Detector mode: INTEGRATING / Number grids imaged: 3 / Number real images: 5500 / Average exposure time: 1.1 sec. / Average electron dose: 32.0 e/Å2 Details: Images were collected in movie-mode at 32 frames per second |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Particle selection | Number selected: 1182309 |
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CTF correction | Software - Name: CTFFIND (ver. 3) |
Startup model | Type of model: PDB ENTRY PDB model - PDB ID: |
Initial angle assignment | Type: NOT APPLICABLE / Software - Name: RELION (ver. 1.4) |
Final 3D classification | Software - Name: RELION (ver. 1.4) |
Final angle assignment | Type: ANGULAR RECONSTITUTION / Software - Name: RELION (ver. 1.4) |
Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 5.75 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 1.4) / Number images used: 12586 |
-Atomic model buiding 1
Refinement | Space: RECIPROCAL / Protocol: OTHER / Target criteria: FSC |
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Output model | PDB-6gsn: |