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Structure paper

TitleEnsemble cryo-EM elucidates the mechanism of translation fidelity.
Journal, issue, pagesNature, Vol. 546, Issue 7656, Page 113-117, Year 2017
Publish dateJun 1, 2017
AuthorsAnna B Loveland / Gabriel Demo / Nikolaus Grigorieff / Andrei A Korostelev /
PubMed AbstractGene translation depends on accurate decoding of mRNA, the structural mechanism of which remains poorly understood. Ribosomes decode mRNA codons by selecting cognate aminoacyl-tRNAs delivered by ...Gene translation depends on accurate decoding of mRNA, the structural mechanism of which remains poorly understood. Ribosomes decode mRNA codons by selecting cognate aminoacyl-tRNAs delivered by elongation factor Tu (EF-Tu). Here we present high-resolution structural ensembles of ribosomes with cognate or near-cognate aminoacyl-tRNAs delivered by EF-Tu. Both cognate and near-cognate tRNA anticodons explore the aminoacyl-tRNA-binding site (A site) of an open 30S subunit, while inactive EF-Tu is separated from the 50S subunit. A transient conformation of decoding-centre nucleotide G530 stabilizes the cognate codon-anticodon helix, initiating step-wise 'latching' of the decoding centre. The resulting closure of the 30S subunit docks EF-Tu at the sarcin-ricin loop of the 50S subunit, activating EF-Tu for GTP hydrolysis and enabling accommodation of the aminoacyl-tRNA. By contrast, near-cognate complexes fail to induce the G530 latch, thus favouring open 30S pre-accommodation intermediates with inactive EF-Tu. This work reveals long-sought structural differences between the pre-accommodation of cognate and near-cognate tRNAs that elucidate the mechanism of accurate decoding.
External linksNature / PubMed:28538735 / PubMed Central
MethodsEM (single particle)
Resolution3.2 - 4.0 Å
Structure data

8615

5uyk

EMDB-8615, PDB-5uyk:
70S ribosome bound with cognate ternary complex not base-paired to A site codon (Structure I)
Method: EM (single particle) / Resolution: 3.9 Å

8616

5uyl

EMDB-8616, PDB-5uyl:
70S ribosome bound with cognate ternary complex base-paired to A site codon (Structure II)
Method: EM (single particle) / Resolution: 3.6 Å

8617

5uym

EMDB-8617, PDB-5uym:
70S ribosome bound with cognate ternary complex base-paired to A site codon, closed 30S (Structure III)
Method: EM (single particle) / Resolution: 3.2 Å

8618

5uyn

EMDB-8618, PDB-5uyn:
70S ribosome bound with near-cognate ternary complex not base-paired to A site codon (Structure I-nc)
Method: EM (single particle) / Resolution: 4.0 Å

8619

5uyp

EMDB-8619, PDB-5uyp:
70S ribosome bound with near-cognate ternary complex base-paired to A site codon, open 30S (Structure II-nc)
Method: EM (single particle) / Resolution: 3.9 Å

8620

5uyq

EMDB-8620, PDB-5uyq:
70S ribosome bound with near-cognate ternary complex base-paired to A site codon, closed 30S (Structure III-nc)
Method: EM (single particle) / Resolution: 3.8 Å

Chemicals

ChemComp-GCP:
PHOSPHOMETHYLPHOSPHONIC ACID GUANYLATE ESTER / GMP-PCP, energy-carrying molecule analogue*YM

ChemComp-FME:
N-FORMYLMETHIONINE

ChemComp-PHE:
PHENYLALANINE

ChemComp-MG:
Unknown entry

ChemComp-ZN:
Unknown entry

ChemComp-LYS:
LYSINE

Source
  • escherichia coli (strain k12) (bacteria)
  • escherichia coli k-12 (bacteria)
KeywordsRIBOSOME / ternary complex / EF-Tu / tRNA

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