Aminoglycoside activity observed on single pre-translocation ribosome complexes. Academic Article uri icon

Overview

MeSH

  • Binding Sites
  • Conserved Sequence
  • Cryoelectron Microscopy
  • Fluorescence Resonance Energy Transfer
  • Kanamycin
  • Kinetics
  • Models, Molecular
  • Mutation
  • Neomycin
  • Oligonucleotides
  • Protein Binding
  • Protein Transport
  • RNA
  • RNA, Transfer
  • Reproducibility of Results

MeSH Major

  • Aminoglycosides
  • Ribosomes

abstract

  • Aminoglycoside-class antibiotics bind directly to ribosomal RNA, imparting pleiotropic effects on ribosome function. Despite in-depth structural investigations of aminoglycoside-RNA oligonucleotide and aminoglycoside-ribosome interactions, mechanisms explaining the unique ribosome inhibition profiles of chemically similar aminoglycosides remain elusive. Here, using single-molecule fluorescence resonance energy transfer (smFRET) methods, we show that high-affinity aminoglycoside binding to the conserved decoding site region of the functional pre-translocation ribosome complex specifically remodels the nature of intrinsic dynamic processes within the particle. The extents of these effects, which are distinct for each member of the aminoglycoside class, strongly correlate with their inhibition of EF-G-catalyzed translocation. Neomycin, a 4,5-linked aminoglycoside, binds with lower affinity to one or more secondary binding sites, mediating distinct structural and dynamic perturbations that further enhance translocation inhibition. These new insights help explain why closely related aminoglycosides elicit pleiotropic translation activities and demonstrate the potential utility of smFRET as a tool for dissecting the mechanisms of antibiotic action.

publication date

  • January 2010

has subject area

  • Aminoglycosides
  • Binding Sites
  • Conserved Sequence
  • Cryoelectron Microscopy
  • Fluorescence Resonance Energy Transfer
  • Kanamycin
  • Kinetics
  • Models, Molecular
  • Mutation
  • Neomycin
  • Oligonucleotides
  • Protein Binding
  • Protein Transport
  • RNA
  • RNA, Transfer
  • Reproducibility of Results
  • Ribosomes

Research

keywords

  • Journal Article

Identity

Language

  • eng

PubMed Central ID

  • PMC2914512

Digital Object Identifier (DOI)

  • 10.1038/nchembio.274

PubMed ID

  • 19946275

Additional Document Info

start page

  • 54

end page

  • 62

volume

  • 6

number

  • 1