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PMID: 28538735 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Ensemble cryo-EM elucidates the mechanism of translation fidelity.

Nature ·Vol. 546 ·No. 7656 ·2017-00-01 ·Pages 113-117

Loveland AB, Demo G, Grigorieff N, Korostelev AA

Abstract

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.

MeSH Terms
Anticodon/chemistry,genetics,ultrastructure Codon/chemistry,genetics,ultrastructure Cryoelectron Microscopy Escherichia coli/chemistry,genetics,ultrastructure GTP Phosphohydrolases/metabolism,ultrastructure Guanosine Triphosphate/metabolism Hydrolysis Models, Molecular Peptide Elongation Factor Tu/metabolism,ultrastructure Protein Biosynthesis Protein Domains RNA, Ribosomal, 16S/genetics,metabolism,ultrastructure RNA, Transfer, Amino Acyl/genetics,metabolism,ultrastructure Ribosome Subunits/chemistry,metabolism,ultrastructure Ribosomes/chemistry,metabolism,ultrastructure
Chemicals
Anticodon Codon RNA, Ribosomal, 16S RNA, Transfer, Amino Acyl Guanosine Triphosphate GTP Phosphohydrolases Peptide Elongation Factor Tu
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Loveland Anna B
RNA Therapeutics Institute, Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 368 Plantation Street, Worcester, Massachusetts 01605, USA.
Demo Gabriel
RNA Therapeutics Institute, Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 368 Plantation Street, Worcester, Massachusetts 01605, USA.
Grigorieff Nikolaus
Janelia Research Campus, Howard Hughes Medical Institute, 19700 Helix Drive, Ashburn, Virginia 20147, USA.
Korostelev Andrei A
RNA Therapeutics Institute, Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 368 Plantation Street, Worcester, Massachusetts 01605, USA.
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Article Info
Journal
Nature
Abbr.
Nature
ISSN
1476-4687
DOI
10.1038/nature22397
Published
2017-00-01
Epub
2017-00-24
Pages
113-117
Language
English
Region
England
NLM ID
0410462
PMCID
PMC5657493
Subset
IM
Grants
NIGMS NIH HHS · P01 GM062580 · United States
NIH HHS · GM107465 · United States
Howard Hughes Medical Institute · United States
NIGMS NIH HHS · R01 GM106105 · United States
NIGMS NIH HHS · R01 GM107465 · United States
External Links
PubMed source DOI: 10.1038/nature22397 PMC Full Text View journal details
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