Multiperspective smFRET reveals rate-determining late intermediates of ribosomal translocation. Academic Article uri icon

Overview

MeSH

  • Fluorescence Resonance Energy Transfer
  • Guanosine Diphosphate
  • Models, Molecular
  • Protein Transport

MeSH Major

  • Escherichia coli
  • Escherichia coli Proteins
  • Peptide Elongation Factor G
  • RNA, Bacterial
  • RNA, Transfer, Amino Acyl
  • Ribosomes

abstract

  • Directional translocation of the ribosome through the mRNA open reading frame is a critical determinant of translational fidelity. This process entails a complex interplay of large-scale conformational changes within the actively translating particle, which together coordinate the movement of tRNA and mRNA substrates with respect to the large and small ribosomal subunits. Using pre-steady state, single-molecule fluorescence resonance energy transfer imaging, we tracked the nature and timing of these conformational events within the Escherichia coli ribosome from five structural perspectives. Our investigations revealed direct evidence of structurally and kinetically distinct late intermediates during substrate movement, whose resolution determines the rate of translocation. These steps involve intramolecular events within the EF-G-GDP-bound ribosome, including exaggerated, reversible fluctuations of the small-subunit head domain, which ultimately facilitate peptidyl-tRNA's movement into its final post-translocation position.

publication date

  • April 2016

has subject area

  • Escherichia coli
  • Escherichia coli Proteins
  • Fluorescence Resonance Energy Transfer
  • Guanosine Diphosphate
  • Models, Molecular
  • Peptide Elongation Factor G
  • Protein Transport
  • RNA, Bacterial
  • RNA, Transfer, Amino Acyl
  • Ribosomes

Research

keywords

  • Journal Article

Identity

Language

  • eng

PubMed Central ID

  • PMC4821728

Digital Object Identifier (DOI)

  • 10.1038/nsmb.3177

PubMed ID

  • 26926435

Additional Document Info

start page

  • 333

end page

  • 341

volume

  • 23

number

  • 4