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Yorodumi- EMDB-43930: 70S initiation complex (tRNA-fMet M1, initiation factor 2 + CUG s... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-43930 | ||||||||||||||||||
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Title | 70S initiation complex (tRNA-fMet M1, initiation factor 2 + CUG start codon) | ||||||||||||||||||
Map data | Masked, sharpened map used for modeling (produced from masked, unsharpened map using PHENIX Autosharpen with sharpening B-factor of -7.3) | ||||||||||||||||||
Sample |
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Keywords | translation initiation / tRNA-fMet M1 / frameshifting / initiation factor 2 / RIBOSOME | ||||||||||||||||||
Function / homology | Function and homology information guanosine tetraphosphate binding / DnaA-L2 complex / negative regulation of DNA-templated DNA replication initiation / translation initiation factor activity / ribosome assembly / assembly of large subunit precursor of preribosome / cytosolic ribosome assembly / mRNA 5'-UTR binding / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding ...guanosine tetraphosphate binding / DnaA-L2 complex / negative regulation of DNA-templated DNA replication initiation / translation initiation factor activity / ribosome assembly / assembly of large subunit precursor of preribosome / cytosolic ribosome assembly / mRNA 5'-UTR binding / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / large ribosomal subunit / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / large ribosomal subunit rRNA binding / transferase activity / cytosolic small ribosomal subunit / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / tRNA binding / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / ribonucleoprotein complex / translation / response to antibiotic / GTPase activity / mRNA binding / GTP binding / RNA binding / zinc ion binding / membrane / cytosol / cytoplasm Similarity search - Function | ||||||||||||||||||
Biological species | Escherichia coli (E. coli) | ||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.6 Å | ||||||||||||||||||
Authors | Mattingly JM / Nguyen HA / Dunham CM | ||||||||||||||||||
Funding support | United States, 5 items
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Citation | Journal: J Biol Chem / Year: 2024 Title: Structural analysis of noncanonical translation initiation complexes. Authors: Jacob M Mattingly / Ha An Nguyen / Bappaditya Roy / Kurt Fredrick / Christine M Dunham / Abstract: Translation initiation is a highly regulated, multi-step process that is critical for efficient and accurate protein synthesis. In bacteria, initiation begins when mRNA, initiation factors, and a ...Translation initiation is a highly regulated, multi-step process that is critical for efficient and accurate protein synthesis. In bacteria, initiation begins when mRNA, initiation factors, and a dedicated initiator fMet-tRNA bind the small (30S) ribosomal subunit. Specific binding of fMet-tRNA in the peptidyl (P) site is mediated by the inspection of the fMet moiety by initiation factor IF2 and of three conserved G-C base pairs in the tRNA anticodon stem by the 30S head domain. Tandem A-minor interactions form between 16S ribosomal RNA nucleotides A1339 and G1338 and tRNA base pairs G30-C40 and G29-C41, respectively. Swapping the G30-C40 pair of tRNA with C-G (called tRNA M1) reduces discrimination against the noncanonical start codon CUG in vitro, suggesting crosstalk between the gripping of the anticodon stem and recognition of the start codon. Here, we solved electron cryomicroscopy structures of Escherichia coli 70S initiation complexes containing the fMet-tRNA M1 variant paired to the noncanonical CUG start codon, in the presence or absence of IF2 and the non-hydrolyzable GTP analog GDPCP, alongside structures of 70S initiation complexes containing this tRNA variant paired to the canonical bacterial start codons AUG, GUG, and UUG. We find that the M1 mutation weakens A-minor interactions between tRNA and 16S nucleotides A1339 and G1338, with IF2 strengthening the interaction of G1338 with the tRNA minor groove. These structures suggest how even slight changes to the recognition of the fMet-tRNA anticodon stem by the ribosome can impact the start codon selection. | ||||||||||||||||||
History |
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-Structure visualization
-Downloads & links
-EMDB archive
Map data | emd_43930.map.gz | 466.9 MB | EMDB map data format | |
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Header (meta data) | emd-43930-v30.xml emd-43930.xml | 75 KB 75 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_43930_fsc.xml | 18.1 KB | Display | FSC data file |
Images | emd_43930.png | 211.2 KB | ||
Masks | emd_43930_msk_1.map | 512 MB | Mask map | |
Filedesc metadata | emd-43930.cif.gz | 14.4 KB | ||
Others | emd_43930_additional_1.map.gz emd_43930_additional_2.map.gz emd_43930_half_map_1.map.gz emd_43930_half_map_2.map.gz | 409.4 MB 477.4 MB 413 MB 412.5 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-43930 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-43930 | HTTPS FTP |
-Validation report
Summary document | emd_43930_validation.pdf.gz | 1.1 MB | Display | EMDB validaton report |
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Full document | emd_43930_full_validation.pdf.gz | 1.1 MB | Display | |
Data in XML | emd_43930_validation.xml.gz | 26.4 KB | Display | |
Data in CIF | emd_43930_validation.cif.gz | 34.7 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-43930 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-43930 | HTTPS FTP |
-Related structure data
Related structure data | 9ax8MC 9ax7C 9cg5C 9cg6C 9cg7C C: citing same article (ref.) M: atomic model generated by this map |
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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_43930.map.gz / Format: CCP4 / Size: 512 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | Masked, sharpened map used for modeling (produced from masked, unsharpened map using PHENIX Autosharpen with sharpening B-factor of -7.3) | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.814 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
File | emd_43930_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
+Entire : E. coli 70S ribosome in complex with fMet-tRNA-fMet M1, initiatio...
+Supramolecule #1: E. coli 70S ribosome in complex with fMet-tRNA-fMet M1, initiatio...
+Macromolecule #1: 50S ribosomal protein L27
+Macromolecule #2: 50S ribosomal protein L28
+Macromolecule #3: 50S ribosomal protein L29
+Macromolecule #4: 50S ribosomal protein L30
+Macromolecule #5: 50S ribosomal protein L3
+Macromolecule #6: 50S ribosomal protein L32
+Macromolecule #7: 50S ribosomal protein L33
+Macromolecule #8: 50S ribosomal protein L34
+Macromolecule #9: 50S ribosomal protein L35
+Macromolecule #10: 50S ribosomal protein L2
+Macromolecule #11: 50S ribosomal protein L4
+Macromolecule #12: 50S ribosomal protein L5
+Macromolecule #13: 50S ribosomal protein L6
+Macromolecule #14: 50S ribosomal protein L13
+Macromolecule #15: 50S ribosomal protein L14
+Macromolecule #16: 50S ribosomal protein L15
+Macromolecule #17: 50S ribosomal protein L16
+Macromolecule #18: 50S ribosomal protein L17
+Macromolecule #19: 50S ribosomal protein L18
+Macromolecule #20: 50S ribosomal protein L19
+Macromolecule #21: 50S ribosomal protein L20
+Macromolecule #22: 50S ribosomal protein L21
+Macromolecule #23: 50S ribosomal protein L22
+Macromolecule #24: 50S ribosomal protein L23
+Macromolecule #25: 50S ribosomal protein L24
+Macromolecule #26: 50S ribosomal protein L25
+Macromolecule #28: 50S ribosomal protein L36
+Macromolecule #30: 30S ribosomal protein S2
+Macromolecule #31: 30S ribosomal protein S3
+Macromolecule #32: 30S ribosomal protein S4
+Macromolecule #33: 30S ribosomal protein S5
+Macromolecule #34: 30S ribosomal protein S6
+Macromolecule #35: 30S ribosomal protein S7
+Macromolecule #36: 30S ribosomal protein S8
+Macromolecule #37: 30S ribosomal protein S9
+Macromolecule #38: 30S ribosomal protein S10
+Macromolecule #39: 30S ribosomal protein S11
+Macromolecule #40: 30S ribosomal protein S12
+Macromolecule #41: 30S ribosomal protein S13
+Macromolecule #42: 30S ribosomal protein S14
+Macromolecule #43: 30S ribosomal protein S15
+Macromolecule #44: 30S ribosomal protein S16
+Macromolecule #45: 30S ribosomal protein S17
+Macromolecule #46: 30S ribosomal protein S18
+Macromolecule #47: 30S ribosomal protein S19
+Macromolecule #48: 30S ribosomal protein S20
+Macromolecule #49: 30S ribosomal protein S21
+Macromolecule #52: Translation initiation factor IF2
+Macromolecule #27: 5S ribosomal RNA
+Macromolecule #29: 16S ribosomal RNA
+Macromolecule #50: mRNA
+Macromolecule #51: P-site tRNA-fMet M1
+Macromolecule #53: 23S ribosomal RNA
+Macromolecule #54: MAGNESIUM ION
+Macromolecule #55: PHOSPHOMETHYLPHOSPHONIC ACID GUANYLATE ESTER
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 |
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Grid | Model: C-flat-1.2/1.3 / Material: GOLD / Mesh: 300 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 15 sec. |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | TFS KRIOS |
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Image recording | Film or detector model: TFS FALCON 4i (4k x 4k) / Average electron dose: 50.0 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: 2.5 µm / Nominal defocus min: 0.5 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |