tRNA selection and kinetic proofreading in translation. Academic Article uri icon

Overview

MeSH

  • Codon
  • Energy Transfer
  • Fluorescence
  • GTP Phosphohydrolases

MeSH Major

  • Protein Biosynthesis
  • RNA, Transfer

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.

publication date

  • October 2004

has subject area

  • Codon
  • Energy Transfer
  • Fluorescence
  • GTP Phosphohydrolases
  • Protein Biosynthesis
  • RNA, Transfer

Research

keywords

  • Journal Article

Identity

Language

  • eng

Digital Object Identifier (DOI)

  • 10.1038/nsmb831

PubMed ID

  • 15448679

Additional Document Info

start page

  • 1008

end page

  • 1014

volume

  • 11

number

  • 10