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Open data
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Basic information
| Entry | Database: EMDB / ID: EMD-4242 | |||||||||
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| Title | Human cap-dependent 48S pre-initiation complex | |||||||||
Map data | human 48S pre-initiation complex, reconstituted in the presence of capped mRNA, eIF4B and eIF4F, displaying eIF2 ternary complex, eIF3 and eIF4B | |||||||||
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Keywords | Translation initiation / 48S complex / capped mRNA / initiation factor 4B / start codon recognition / RIBOSOME | |||||||||
| Function / homology | Function and homology informationeukaryotic translation initiation factor 4F complex assembly / RNA strand-exchange activity / Cellular response to mitochondrial stress / viral translational termination-reinitiation / Response of EIF2AK1 (HRI) to heme deficiency / Recycling of eIF2:GDP / methionyl-initiator methionine tRNA binding / Activation of the mRNA upon binding of the cap-binding complex and eIFs, and subsequent binding to 43S / eukaryotic translation initiation factor 3 complex, eIF3e / eukaryotic translation initiation factor 3 complex, eIF3m ...eukaryotic translation initiation factor 4F complex assembly / RNA strand-exchange activity / Cellular response to mitochondrial stress / viral translational termination-reinitiation / Response of EIF2AK1 (HRI) to heme deficiency / Recycling of eIF2:GDP / methionyl-initiator methionine tRNA binding / Activation of the mRNA upon binding of the cap-binding complex and eIFs, and subsequent binding to 43S / eukaryotic translation initiation factor 3 complex, eIF3e / eukaryotic translation initiation factor 3 complex, eIF3m / RNA strand annealing activity / PERK regulates gene expression / eukaryotic translation initiation factor 2 complex / eukaryotic translation initiation factor 4F complex / Z-decay: degradation of maternal mRNAs by zygotically expressed factors / formation of cytoplasmic translation initiation complex / eukaryotic translation initiation factor 3 complex / eukaryotic 43S preinitiation complex / translation factor activity, RNA binding / Deadenylation of mRNA / formation of translation preinitiation complex / eukaryotic 48S preinitiation complex / M-decay: degradation of maternal mRNAs by maternally stored factors / negative regulation of endoplasmic reticulum unfolded protein response / oxidized pyrimidine DNA binding / response to TNF agonist / positive regulation of base-excision repair / positive regulation of gastrulation / positive regulation of intrinsic apoptotic signaling pathway in response to DNA damage / protein tyrosine kinase inhibitor activity / regulation of translational initiation / IRE1-RACK1-PP2A complex / positive regulation of Golgi to plasma membrane protein transport / protein-synthesizing GTPase / positive regulation of DNA-templated transcription initiation / TNFR1-mediated ceramide production / nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / negative regulation of RNA splicing / supercoiled DNA binding / neural crest cell differentiation / NF-kappaB complex / negative regulation of DNA repair / oxidized purine DNA binding / cytoplasmic translational initiation / cysteine-type endopeptidase activator activity involved in apoptotic process / regulation of establishment of cell polarity / negative regulation of intrinsic apoptotic signaling pathway in response to hydrogen peroxide / negative regulation of bicellular tight junction assembly / ubiquitin-like protein conjugating enzyme binding / rRNA modification in the nucleus and cytosol / negative regulation of phagocytosis / erythrocyte homeostasis / Formation of the ternary complex, and subsequently, the 43S complex / cytoplasmic side of rough endoplasmic reticulum membrane / protein kinase A binding / ion channel inhibitor activity / Ribosomal scanning and start codon recognition / laminin receptor activity / pigmentation / Translation initiation complex formation / positive regulation of mitochondrial depolarization / negative regulation of Wnt signaling pathway / BH3 domain binding / negative regulation of translational frameshifting / TOR signaling / Protein hydroxylation / SARS-CoV-1 modulates host translation machinery / regulation of adenylate cyclase-activating G protein-coupled receptor signaling pathway / iron-sulfur cluster binding / mTORC1-mediated signalling / positive regulation of GTPase activity / ribosomal small subunit binding / Peptide chain elongation / regulation of cell division / Selenocysteine synthesis / cellular response to ethanol / Formation of a pool of free 40S subunits / positive regulation of intrinsic apoptotic signaling pathway by p53 class mediator / Eukaryotic Translation Termination / protein serine/threonine kinase inhibitor activity / SRP-dependent cotranslational protein targeting to membrane / Response of EIF2AK4 (GCN2) to amino acid deficiency / negative regulation of protein binding / Viral mRNA Translation / ubiquitin ligase inhibitor activity / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / positive regulation of signal transduction by p53 class mediator / GTP hydrolysis and joining of the 60S ribosomal subunit / negative regulation of ubiquitin-dependent protein catabolic process / L13a-mediated translational silencing of Ceruloplasmin expression / Major pathway of rRNA processing in the nucleolus and cytosol / positive regulation of microtubule polymerization / regulation of translational fidelity / phagocytic cup / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / positive regulation of intrinsic apoptotic signaling pathway / spindle assembly / Nuclear events stimulated by ALK signaling in cancer / translation regulator activity / rough endoplasmic reticulum Similarity search - Function | |||||||||
| Biological species | Homo sapiens (human) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 6.3 Å | |||||||||
Authors | Schaffitzel C | |||||||||
| Funding support | Belgium, 2 items
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Citation | Journal: Nucleic Acids Res / Year: 2018Title: Structure of a human cap-dependent 48S translation pre-initiation complex. Authors: Boris Eliseev / Lahari Yeramala / Alexander Leitner / Manikandan Karuppasamy / Etienne Raimondeau / Karine Huard / Elena Alkalaeva / Ruedi Aebersold / Christiane Schaffitzel / ![]() Abstract: Eukaryotic translation initiation is tightly regulated, requiring a set of conserved initiation factors (eIFs). Translation of a capped mRNA depends on the trimeric eIF4F complex and eIF4B to load ...Eukaryotic translation initiation is tightly regulated, requiring a set of conserved initiation factors (eIFs). Translation of a capped mRNA depends on the trimeric eIF4F complex and eIF4B to load the mRNA onto the 43S pre-initiation complex comprising 40S and initiation factors 1, 1A, 2, 3 and 5 as well as initiator-tRNA. Binding of the mRNA is followed by mRNA scanning in the 48S pre-initiation complex, until a start codon is recognised. Here, we use a reconstituted system to prepare human 48S complexes assembled on capped mRNA in the presence of eIF4B and eIF4F. The highly purified h-48S complexes are used for cross-linking/mass spectrometry, revealing the protein interaction network in this complex. We report the electron cryo-microscopy structure of the h-48S complex at 6.3 Å resolution. While the majority of eIF4B and eIF4F appear to be flexible with respect to the ribosome, additional density is detected at the entrance of the 40S mRNA channel which we attribute to the RNA-recognition motif of eIF4B. The eight core subunits of eIF3 are bound at the 40S solvent-exposed side, as well as the subunits eIF3d, eIF3b and eIF3i. elF2 and initiator-tRNA bound to the start codon are present at the 40S intersubunit side. This cryo-EM structure represents a molecular snap-shot revealing the h-48S complex following start codon recognition. | |||||||||
| History |
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Structure visualization
| Movie |
Movie viewer |
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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_4242.map.gz | 32.1 MB | EMDB map data format | |
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| Header (meta data) | emd-4242-v30.xml emd-4242.xml | 74.1 KB 74.1 KB | Display Display | EMDB header |
| Images | emd_4242.png | 56.4 KB | ||
| Filedesc metadata | emd-4242.cif.gz | 16.8 KB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-4242 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-4242 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 6fecMC ![]() 4265C M: atomic model generated by this map C: citing same article ( |
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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_4242.map.gz / Format: CCP4 / Size: 34.3 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Annotation | human 48S pre-initiation complex, reconstituted in the presence of capped mRNA, eIF4B and eIF4F, displaying eIF2 ternary complex, eIF3 and eIF4B | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 2.5 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
+Entire : human cap-dependent 48S translation pre-initiation complex
+Supramolecule #1: human cap-dependent 48S translation pre-initiation complex
+Supramolecule #2: human cap-dependent 48S translation pre-initiation complex
+Supramolecule #3: mRNA
+Supramolecule #4: Eukaryotic translation initiation factor 4B
+Macromolecule #1: Eukaryotic translation initiation factor 3 subunit A
+Macromolecule #2: Eukaryotic translation initiation factor 3 subunit C
+Macromolecule #3: Eukaryotic translation initiation factor 3 subunit E
+Macromolecule #4: Eukaryotic translation initiation factor 3 subunit F
+Macromolecule #5: Eukaryotic translation initiation factor 3 subunit H
+Macromolecule #6: Eukaryotic translation initiation factor 3 subunit K
+Macromolecule #7: Eukaryotic translation initiation factor 3 subunit L
+Macromolecule #8: Eukaryotic translation initiation factor 3 subunit M
+Macromolecule #9: EUKARYOTIC TRANSLATION INITIATION FACTOR 3 SUBUNIT D
+Macromolecule #12: 40S ribosomal protein S11
+Macromolecule #13: 40S ribosomal protein S16
+Macromolecule #14: 40S ribosomal protein S4, X isoform
+Macromolecule #15: 40S ribosomal protein S29
+Macromolecule #16: 40S ribosomal protein S9
+Macromolecule #17: 40S ribosomal protein S18
+Macromolecule #19: Eukaryotic translation initiation factor 2 subunit 1
+Macromolecule #20: 40S ribosomal protein S23
+Macromolecule #21: 40S ribosomal protein S19
+Macromolecule #22: Eukaryotic translation initiation factor 2 subunit 3
+Macromolecule #23: 40S ribosomal protein S5
+Macromolecule #24: 40S ribosomal protein S30
+Macromolecule #25: 40S ribosomal protein S25
+Macromolecule #26: 40S ribosomal protein S7
+Macromolecule #27: 40S ribosomal protein S27
+Macromolecule #28: 40S ribosomal protein S13
+Macromolecule #29: 40S ribosomal protein S15a
+Macromolecule #30: 40S ribosomal protein S21
+Macromolecule #31: 40S ribosomal protein S2
+Macromolecule #32: EUKARYOTIC TRANSLATION INITIATION FACTOR 2 BETA SUBUNIT (eIF2-Beta)
+Macromolecule #33: 40S ribosomal protein S17
+Macromolecule #34: 40S ribosomal protein SA
+Macromolecule #35: 40S ribosomal protein S3
+Macromolecule #36: 40S ribosomal protein S20
+Macromolecule #37: 40S ribosomal protein S3a
+Macromolecule #38: 40S ribosomal protein S14
+Macromolecule #39: 40S ribosomal protein S26
+Macromolecule #40: 40S ribosomal protein S28
+Macromolecule #41: Receptor of activated protein C kinase 1
+Macromolecule #42: 40S ribosomal protein S15
+Macromolecule #43: 40S ribosomal protein S8
+Macromolecule #44: Ubiquitin-40S ribosomal protein S27a
+Macromolecule #45: 40S ribosomal protein S6
+Macromolecule #46: 40S ribosomal protein S12
+Macromolecule #47: 40S ribosomal protein S24
+Macromolecule #48: 40S ribosomal protein S10
+Macromolecule #49: Eukaryotic translation initiation factor 4B
+Macromolecule #50: Eukaryotic translation initiation factor 3 subunit B
+Macromolecule #10: 18S ribosomal RNA
+Macromolecule #11: Messenger RNA (26-MER)
+Macromolecule #18: Transfer RNA (75-MER)
+Macromolecule #51: water
-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 Details: 20 mM Tris HCl, 50 mM KOAc, 2.5 mM MgCl2, 2 mM DTT, 0.25 mM spermidine 0.25 mM GMPPNP |
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| Grid | Model: Quantifoil R2/2 |
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Image recording | Film or detector model: FEI FALCON II (4k x 4k) / Detector mode: INTEGRATING / Average electron dose: 30.0 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 4.0 µm / Nominal defocus min: 1.5 µm / Nominal magnification: 112000 |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
| Refinement | Space: REAL / Protocol: RIGID BODY FIT / Target criteria: Cross-correlation coefficient |
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| Output model | ![]() PDB-6fec: |
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About Yorodumi



Keywords
Homo sapiens (human)
Authors
Belgium, 2 items
Citation

UCSF Chimera







































Z (Sec.)
Y (Row.)
X (Col.)

























