+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-17330 | |||||||||
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Title | 48S late-stage initiation complex with non methylated mRNA | |||||||||
Map data | ||||||||||
Sample |
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Keywords | translation initiation / ribosome / non methylated mRNA / TRANSLATION | |||||||||
Function / homology | Function and homology information Translation initiation complex formation / Formation of the ternary complex, and subsequently, the 43S complex / Ribosomal scanning and start codon recognition / Major pathway of rRNA processing in the nucleolus and cytosol / GTP hydrolysis and joining of the 60S ribosomal subunit / 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) ...Translation initiation complex formation / Formation of the ternary complex, and subsequently, the 43S complex / Ribosomal scanning and start codon recognition / Major pathway of rRNA processing in the nucleolus and cytosol / GTP hydrolysis and joining of the 60S ribosomal subunit / 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 2 complex / ribosomal subunit / eukaryotic 48S preinitiation complex / laminin receptor activity / positive regulation of signal transduction by p53 class mediator / ubiquitin ligase inhibitor activity / phagocytic cup / ribosomal small subunit binding / 90S preribosome / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / regulation of translational fidelity / ribosomal subunit export from nucleus / ribosomal small subunit export from nucleus / translation regulator activity / translational termination / laminin binding / rough endoplasmic reticulum / 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) / gastrulation / MDM2/MDM4 family protein binding / cytosolic ribosome / translation initiation factor activity / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / class I DNA-(apurinic or apyrimidinic site) endonuclease activity / DNA-(apurinic or apyrimidinic site) lyase / rescue of stalled ribosome / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / cellular response to leukemia inhibitory factor / small-subunit processome / translational initiation / protein kinase C binding / positive regulation of apoptotic signaling pathway / positive regulation of protein-containing complex assembly / spindle / modification-dependent protein catabolic process / cytoplasmic stress granule / protein tag activity / rRNA processing / ribosomal small subunit biogenesis / rhythmic process / positive regulation of canonical Wnt signaling pathway / small ribosomal subunit rRNA binding / ribosome binding / regulation of translation / ribosomal small subunit assembly / virus receptor activity / small ribosomal subunit / cytosolic small ribosomal subunit / cytoplasmic translation / perikaryon / cytosolic large ribosomal subunit / mitochondrial inner membrane / postsynaptic density / cell differentiation / rRNA binding / ribosome / protein ubiquitination / structural constituent of ribosome / iron ion binding / positive regulation of protein phosphorylation / ribonucleoprotein complex / translation / cell division / DNA repair / mRNA binding / centrosome / dendrite / ubiquitin protein ligase binding / synapse / nucleolus / apoptotic process / perinuclear region of cytoplasm / Golgi apparatus / ATP hydrolysis activity / DNA binding / RNA binding / zinc ion binding / ATP binding / membrane / nucleus / metal ion binding / plasma membrane / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Oryctolagus cuniculus (rabbit) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.3 Å | |||||||||
Authors | Guca E / Lima LHF / Boissier F / Hashem Y | |||||||||
Funding support | European Union, 1 items
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Citation | Journal: Mol Cell / Year: 2024 Title: N-methyladenosine in 5' UTR does not promote translation initiation. Authors: Ewelina Guca / Rodrigo Alarcon / Michael Z Palo / Leonardo Santos / Santiago Alonso-Gil / Marcos Davyt / Leonardo H F de Lima / Fanny Boissier / Sarada Das / Bojan Zagrovic / Joseph D ...Authors: Ewelina Guca / Rodrigo Alarcon / Michael Z Palo / Leonardo Santos / Santiago Alonso-Gil / Marcos Davyt / Leonardo H F de Lima / Fanny Boissier / Sarada Das / Bojan Zagrovic / Joseph D Puglisi / Yaser Hashem / Zoya Ignatova / Abstract: The most abundant N-methyladenosine (mA) modification on mRNAs is installed non-stoichiometrically across transcripts, with 5' untranslated regions (5' UTRs) being the least conductive. 5' UTRs are ...The most abundant N-methyladenosine (mA) modification on mRNAs is installed non-stoichiometrically across transcripts, with 5' untranslated regions (5' UTRs) being the least conductive. 5' UTRs are essential for translation initiation, yet the molecular mechanisms orchestrated by mA remain poorly understood. Here, we combined structural, biochemical, and single-molecule approaches and show that at the most common position, a single mA does not affect translation yields, the kinetics of translation initiation complex assembly, or start codon recognition both under permissive growth and following exposure to oxidative stress. Cryoelectron microscopy (cryo-EM) structures of the late preinitiation complex reveal that mA purine ring established stacking interactions with an arginine side chain of the initiation factor eIF2α, although with only a marginal energy contribution, as estimated computationally. These findings provide molecular insights into mA interactions with the initiation complex and suggest that the subtle stabilization is unlikely to affect the translation dynamics under homeostatic conditions or stress. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_17330.map.gz | 170.6 MB | EMDB map data format | |
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Header (meta data) | emd-17330-v30.xml emd-17330.xml | 56.4 KB 56.4 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_17330_fsc.xml | 14.2 KB | Display | FSC data file |
Images | emd_17330.png | 32.1 KB | ||
Filedesc metadata | emd-17330.cif.gz | 12.1 KB | ||
Others | emd_17330_additional_1.map.gz emd_17330_half_map_1.map.gz emd_17330_half_map_2.map.gz | 190.8 MB 168.3 MB 168.3 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-17330 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-17330 | HTTPS FTP |
-Validation report
Summary document | emd_17330_validation.pdf.gz | 1.1 MB | Display | EMDB validaton report |
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Full document | emd_17330_full_validation.pdf.gz | 1.1 MB | Display | |
Data in XML | emd_17330_validation.xml.gz | 21.8 KB | Display | |
Data in CIF | emd_17330_validation.cif.gz | 28.4 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-17330 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-17330 | HTTPS FTP |
-Related structure data
Related structure data | 8p09MC 8p03C M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_17330.map.gz / Format: CCP4 / Size: 216 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.1 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Additional map: #1
File | emd_17330_additional_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_17330_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_17330_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
+Entire : 48S late-stage initiation complex with non methylated mRNA
+Supramolecule #1: 48S late-stage initiation complex with non methylated mRNA
+Macromolecule #1: initiator methionylated tRNA
+Macromolecule #2: 18S ribosomal RNA
+Macromolecule #3: mRNA
+Macromolecule #4: Eukaryotic translation initiation factor 2 subunit 1
+Macromolecule #5: 40S ribosomal protein SA
+Macromolecule #6: ribosomal protein eS1
+Macromolecule #7: 40S ribosomal protein S2
+Macromolecule #8: Ribosomal protein S3
+Macromolecule #9: 40S ribosomal protein S4
+Macromolecule #10: Ribosomal protein S5
+Macromolecule #11: 40S ribosomal protein S6
+Macromolecule #12: ribosomal protein eS7
+Macromolecule #13: 40S ribosomal protein S8
+Macromolecule #14: 40S ribosomal protein S9
+Macromolecule #15: 40S ribosomal protein eS10
+Macromolecule #16: 40S ribosomal protein S11
+Macromolecule #17: 40S ribosomal protein S12
+Macromolecule #18: ribosomal protein uS15
+Macromolecule #19: 40S ribosomal protein uS11
+Macromolecule #20: 40S ribosomal protein uS19
+Macromolecule #21: 40S ribosomal protein uS9
+Macromolecule #22: 40S ribosomal protein eS17
+Macromolecule #23: 40S ribosomal protein uS13
+Macromolecule #24: 40S ribosomal protein eS19
+Macromolecule #25: 40S ribosomal protein uS10
+Macromolecule #26: 40S ribosomal protein S21
+Macromolecule #27: Ribosomal protein S15a
+Macromolecule #28: 40S ribosomal protein S23
+Macromolecule #29: 40S ribosomal protein S24
+Macromolecule #30: 40S ribosomal protein S26
+Macromolecule #31: 40S ribosomal protein S27
+Macromolecule #32: 40S ribosomal protein S28
+Macromolecule #33: 40S ribosomal protein S29
+Macromolecule #34: ribosomal protein eS31
+Macromolecule #35: Ribosomal protein RACK1
+Macromolecule #36: 40S ribosomal protein S30
+Macromolecule #37: Eukaryotic translation initiation factor 4C
+Macromolecule #38: ATP binding cassette subfamily E member 1
+Macromolecule #39: 60S ribosomal protein L41
+Macromolecule #40: 40S ribosomal protein S25
+Macromolecule #41: MAGNESIUM ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.4 |
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Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 4 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TALOS ARCTICA |
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Image recording | Film or detector model: GATAN K2 QUANTUM (4k x 4k) / Average electron dose: 45.0 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: OTHER / Nominal defocus max: 2.3000000000000003 µm / Nominal defocus min: 0.3 µm |
Experimental equipment | Model: Talos Arctica / Image courtesy: FEI Company |