Cited by 466RNA and protein synthesis mechanismsRNA modifications and cancerDNA and Nucleic Acid Chemistry

tRNA selection and kinetic proofreading in translation

Blanchard, Scott C, Gonzalez, Ruben L, Kim, Harold D, Chu, Steven, Puglisi, Joseph D·2004·Nature Structural & Molecular Biology

Abstract

Using single-molecule methods we observed the stepwise movement of aminoacyl-tRNA (aa-tRNA) into the ribosome during selection and kinetic proofreading using single-molecule fluorescence resonance energy transfer (smFRET). Intermediate states in the pathway of tRNA delivery were observed using antibiotics and nonhydrolyzable GTP analogs. We identified three unambiguous FRET states corresponding to initial codon recognition, GTPase-activated and fully accommodated states. The antibiotic tetracycline blocks progression of aa-tRNA from the initial codon recognition state, whereas cleavage of the sarcin-ricin loop impedes progression from the GTPase-activated state. Our data support a model in which ribosomal recognition of correct codon-anticodon pairs drives rotational movement of the incoming complex of EF-Tu-GTP-aa-tRNA toward peptidyl-tRNA during selection on the ribosome. We propose a mechanistic model of initial selection and proofreading.

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Crossref · 10.1038/nsmb831

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Authors
Blanchard, S. C., Gonzalez, R. L., Kim, H. D., Chu, S., & Puglisi, J. D.
Affiliations
  • Stanford University (US)
  • Stanford Medicine (US)
Corresponding author
  • Chu, Steven
  • Puglisi, Joseph D
Venue
Nature Structural & Molecular Biology
Published
2004-09-26
Volume
11
Issue
10
Pages
1008-1014
ISSN
  • 1545-9985 (ISSN-L)
  • 1545-9993
OpenAlex
W2124081058
Cited by
466
References
47
Citations by year
2026: 6 · 2025: 6 · 2024: 16 · 2023: 10 · 2022: 18 · 2021: 11 · 2020: 16 · 2019: 13 · 2018: 20 · 2017: 21 · 2016: 27 · 2015: 18 · 2014: 22 · 2013: 33 · 2012: 26
FWCI
7.82
Type
journal article
Language
en
Open access
closed
Locations
  • Nature Structural & Molecular Biology (publishedVersion)
  • PubMed (publishedVersion)
Funders
National Institute of General Medical Sciences (R01GM051266, GM51266)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA modifications and cancer
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
Keywords
tRNA selection, kinetic proofreading, translation initiation, ribosome, aminoacyl-tRNA, codon-anticodon recognition, sarcin-ricin loop, tetracycline, nonhydrolyzable GTP analogs, single-molecule FRET, ribosome recycling
MeSH terms
Codon, Energy Transfer, Fluorescence, RNA, Transfer, Protein Biosynthesis, GTP Phosphohydrolases

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