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Yorodumi- PDB-8g7p: Structure of the Escherichia coli 70S ribosome in complex with EF... -
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-Basic information
Entry | Database: PDB / ID: 8g7p | |||||||||
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Title | Structure of the Escherichia coli 70S ribosome in complex with EF-Tu and Ile-tRNAIle(LAU) bound to the cognate AUA codon (Structure I) | |||||||||
Components |
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Keywords | RIBOSOME / Elongation factor Tu / lysidine 34 / cryo-EM / tRNA | |||||||||
Function / homology | Function and homology information negative regulation of cytoplasmic translational initiation / guanosine tetraphosphate binding / stringent response / positive regulation of ribosome biogenesis / translation elongation factor activity / translational termination / DnaA-L2 complex / negative regulation of DNA-templated DNA replication initiation / negative regulation of translational initiation / mRNA regulatory element binding translation repressor activity ...negative regulation of cytoplasmic translational initiation / guanosine tetraphosphate binding / stringent response / positive regulation of ribosome biogenesis / translation elongation factor activity / translational termination / DnaA-L2 complex / negative regulation of DNA-templated DNA replication initiation / negative regulation of translational initiation / mRNA regulatory element binding translation repressor activity / ribosome assembly / assembly of large subunit precursor of preribosome / cytosolic ribosome assembly / transcription antitermination / regulation of cell growth / translational initiation / DNA-templated transcription termination / maintenance of translational fidelity / mRNA 5'-UTR binding / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / large ribosomal subunit / ribosome binding / 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) Escherichia phage T4 (virus) | |||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.9 Å | |||||||||
Authors | Rybak, M.Y. / Gagnon, M.G. | |||||||||
Funding support | United States, 2items
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Citation | Journal: Nat Struct Mol Biol / Year: 2024 Title: Structures of the ribosome bound to EF-Tu-isoleucine tRNA elucidate the mechanism of AUG avoidance. Authors: Mariia Yu Rybak / Matthieu G Gagnon / Abstract: The frequency of errors upon decoding of messenger RNA by the bacterial ribosome is low, with one misreading event per 1 × 10 codons. In the universal genetic code, the AUN codon box specifies ...The frequency of errors upon decoding of messenger RNA by the bacterial ribosome is low, with one misreading event per 1 × 10 codons. In the universal genetic code, the AUN codon box specifies two amino acids, isoleucine and methionine. In bacteria and archaea, decoding specificity of the AUA and AUG codons relies on the wobble avoidance strategy that requires modification of C34 in the anticodon loop of isoleucine transfer RNA (tRNA). Bacterial tRNA with 2-lysylcytidine (lysidine) at the wobble position deciphers AUA while avoiding AUG. Here we report cryo-electron microscopy structures of the Escherichia coli 70S ribosome complexed with elongation factor thermo unstable (EF-Tu) and isoleucine-tRNA in the process of decoding AUA and AUG. Lysidine in tRNA excludes AUG by promoting the formation of an unusual Hoogsteen purine-pyrimidine nucleobase geometry at the third position of the codon, weakening the interactions with the mRNA and destabilizing the EF-Tu ternary complex. Our findings elucidate the molecular mechanism by which tRNA specifically decodes AUA over AUG. | |||||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 8g7p.cif.gz | 3.3 MB | Display | PDBx/mmCIF format |
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PDB format | pdb8g7p.ent.gz | Display | PDB format | |
PDBx/mmJSON format | 8g7p.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 8g7p_validation.pdf.gz | 1.9 MB | Display | wwPDB validaton report |
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Full document | 8g7p_full_validation.pdf.gz | 1.9 MB | Display | |
Data in XML | 8g7p_validation.xml.gz | 224.6 KB | Display | |
Data in CIF | 8g7p_validation.cif.gz | 397.6 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/g7/8g7p ftp://data.pdbj.org/pub/pdb/validation_reports/g7/8g7p | HTTPS FTP |
-Related structure data
Related structure data | 29819MC 8g7qC 8g7rC 8g7sC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
-RNA chain , 6 types, 7 molecules awyxvAB
#1: RNA chain | Mass: 499873.406 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: GenBank: 1758835854 | ||||||||
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#22: RNA chain | Mass: 24884.094 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Details: Aminoacylated isoleucine-tRNA / Source: (gene. exp.) Escherichia coli (E. coli) / Strain: MRE600 / Plasmid: pBSTNAV-tRNAIle / Production host: Escherichia coli HB101 (bacteria) #23: RNA chain | | Mass: 24846.902 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: Deacylated Initiator Methionyl-tRNA / Source: (gene. exp.) Escherichia coli (E. coli) / Strain: MRE600 / Plasmid: pBS-tRNAfMet / Production host: Escherichia coli HB101 (bacteria) / References: GenBank: 732678099 #24: RNA chain | | Mass: 8878.456 Da / Num. of mol.: 1 / Source method: obtained synthetically / Details: Synthetic 27-nt M-I mRNA / Source: (synth.) Escherichia phage T4 (virus) #26: RNA chain | | Mass: 941811.562 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 #27: RNA chain | | Mass: 38814.113 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: GenBank: 1817378070 |
-30S ribosomal protein ... , 20 types, 20 molecules bcdefghijklmnopqrstu
#2: Protein | Mass: 26795.695 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: J7QM75 |
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#3: Protein | Mass: 26031.316 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: B7MCS9 |
#4: Protein | Mass: 23514.199 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: P0A7V8 |
#5: Protein | Mass: 17629.398 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: P0A7W1 |
#6: Protein | Mass: 15211.058 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: C3SFQ7 |
#7: Protein | Mass: 17637.445 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: P02359 |
#8: Protein | Mass: 14146.557 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: D7XKZ3 |
#9: Protein | Mass: 14886.270 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: A0A1X3LT86 |
#10: Protein | Mass: 11755.597 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: V0ANK5 |
#11: Protein | Mass: 13871.959 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: A0A0H3PWX2 |
#12: Protein | Mass: 13814.249 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: A0A0F1AUC4 |
#13: Protein | Mass: 13128.467 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: A0A7U9IV78 |
#14: Protein | Mass: 11606.560 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: U9Y6H3 |
#15: Protein | Mass: 10290.816 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: D8EB41 |
#16: Protein | Mass: 9207.572 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: C3SYP2 |
#17: Protein | Mass: 9724.491 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: A0A8E0MHL6 |
#18: Protein | Mass: 9005.472 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: A0A0E2KXL3 |
#19: Protein | Mass: 10455.355 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: S1EA57 |
#20: Protein | Mass: 9708.464 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: C3TRH7 |
#21: Protein | Mass: 8524.039 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: A0A0E2L2J1 |
-Protein , 2 types, 2 molecules zT
#25: Protein | Mass: 43152.219 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: Elongation factor Tu / Source: (gene. exp.) Escherichia coli (E. coli) / Strain: MRE600 Gene: tufA_2, tuf, tufA, tufA_1, tufA_3, tufA_4, tufB, tufB_1, tufB_2, A5U30_004581, A9X72_23960, AAS29_002155, AAS29_004093, ACN68_05365, AM464_13225, AT845_003992, AWP47_16755, B6R15_000408, BANRA_ ...Gene: tufA_2, tuf, tufA, tufA_1, tufA_3, tufA_4, tufB, tufB_1, tufB_2, A5U30_004581, A9X72_23960, AAS29_002155, AAS29_004093, ACN68_05365, AM464_13225, AT845_003992, AWP47_16755, B6R15_000408, BANRA_00548, BANRA_05052, BG944_004744, BGM66_002194, BJJ90_25160, BLM69_004391, BMT49_21045, BMT50_08130, BMT91_03325, BTQ06_27310, BvCmsKKP036_03588, BVL39_05405, BZL69_05770, C0P57_003010, C3F40_15505, C9E67_28700, CA593_06065, CCV12_004444, CDC27_21515, CO706_19465, CR538_24255, CR539_01630, CR628_004228, CV83915_02073, CY655_25255, D1H34_004550, D3Y67_20900, D9E49_24235, DAH19_24560, DAH21_23365, DAH22_22865, DAH27_25225, DAH28_23840, DAH32_23225, DAH33_24610, DAH34_21600, DAH37_22525, DAH38_23185, DAH40_23380, DAH41_23505, DAH42_24340, DAH43_24835, DAH45_24780, DAH46_24615, DAH47_25180, DAH48_24880, DAH49_24790, DNQ45_05140, DS732_01505, DU321_23465, E3N34_20940, E5P27_19115, E5P28_08865, E5P29_14360, EC95NR1_03445, EKI52_13985, EL79_4415, ELT25_22120, ELU85_23630, ELU98_01540, ELV02_23595, ELV12_15580, ExPECSC038_04335, FA849_13360, FA849_22895, FE584_21060, FE587_22575, FEL34_21300, FHO90_24295, FOI11_015750, FOI11_24390, FZN31_22945, FZU14_22815, G9448_13515, GAJ12_23515, GJ11_25160, H0O37_04690, H6Y26_004587, HV109_22460, HV146_01010, HV146_21790, HV209_21230, HVV39_13100, HVW04_13870, HVW43_15020, HVX31_01850, HVX31_19145, HVX32_01070, HVX32_19165, HVY77_24410, HVZ29_01065, HVZ29_19215, HVZ30_01935, HVZ30_19855, HVZ71_23905, HX136_23675, I6H00_17205, I6H01_14295, I6H02_15680, JE86ST02C_45170, JE86ST05C_45270, JFD_05085, JNP96_01240, NCTC10082_02089, NCTC10089_04881, NCTC10418_07157, NCTC10764_02980, NCTC10767_00724, NCTC10865_05903, NCTC11112_02273, NCTC11126_05444, NCTC11181_01973, NCTC11341_02766, NCTC12950_05222, NCTC13127_06172, NCTC13148_04399, NCTC13216_02099, NCTC4450_01743, NCTC7927_05257, NCTC7928_03622, NCTC8333_05566, NCTC8500_05333, NCTC8621_04880, NCTC8959_04056, NCTC8960_02348, NCTC9036_04657, NCTC9037_04790, NCTC9044_02296, NCTC9045_05550, NCTC9073_00863, NCTC9075_06435, NCTC9077_05944, NCTC9111_04963, NCTC9117_05912, NCTC9702_05647, NCTC9706_02011, NCTC9775_03134, NCTC9777_01126, RG28_25305, SAMEA3472044_04882, SAMEA3751407_05016, SAMEA3752557_04835, SAMEA3753106_04223, TUM18780_41760, WP4S18E07_42760 Plasmid: pET21a-EF-Tu / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: E2QJ06 |
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#43: Protein | Mass: 11586.374 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Escherichia coli (E. coli) / Strain: MRE600 / References: UniProt: A0A829CSJ4 |
+50S ribosomal protein ... , 29 types, 29 molecules CDEFGHJLMNOPQRSUVWXYZ12345678
-Non-polymers , 6 types, 1599 molecules
#58: Chemical | ChemComp-MG / #59: Chemical | #60: Chemical | #61: Chemical | ChemComp-GCP / | #62: Chemical | #63: Water | ChemComp-HOH / | |
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-Details
Has ligand of interest | Y |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: Structure of the Escherichia coli 70S ribosome in complex with EF-Tu and Ile-tRNAIle(LAU) bound to the cognate AUA codon (Structure I) Type: RIBOSOME / Entity ID: #1-#57 / Source: NATURAL |
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Molecular weight | Value: 2.6 MDa / Experimental value: NO |
Source (natural) | Organism: Escherichia coli (E. coli) |
Buffer solution | pH: 7.4 |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Specimen support | Grid material: GOLD / Grid mesh size: 200 divisions/in. / Grid type: Quantifoil R2/1 |
Vitrification | Instrument: LEICA EM GP / Cryogen name: ETHANE / Humidity: 85 % / Chamber temperature: 295 K |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: TFS KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 96000 X / Nominal defocus max: 2300 nm / Nominal defocus min: 1000 nm / Cs: 2.7 mm / Alignment procedure: COMA FREE |
Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Average exposure time: 1 sec. / Electron dose: 40 e/Å2 / Detector mode: INTEGRATING / Film or detector model: FEI FALCON III (4k x 4k) / Num. of grids imaged: 1 / Num. of real images: 10381 |
-Processing
EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||||||||||
3D reconstruction | Resolution: 2.9 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 135882 / Symmetry type: POINT | ||||||||||||||||||||||||||||||||
Atomic model building | Protocol: RIGID BODY FIT / Space: REAL | ||||||||||||||||||||||||||||||||
Atomic model building |
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Refine LS restraints |
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