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Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses
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Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses

Published on: February 25, 2011

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Drug sensing by the ribosome induces translational arrest via active site perturbation

Stefan Arenz1, Sezen Meydan2, Agata L Starosta1

  • 1Gene Center and Department for Biochemistry, University of Munich, Feodor-Lynenstr. 25, 81377 Munich, Germany.

Molecular Cell
|October 13, 2014
PubMed

Insights

The ErmCL leader peptide stalls bacterial ribosomes in the presence of erythromycin, preventing protein synthesis. This structural study reveals how the peptide senses the antibiotic to halt translation.

Area of Science:

  • Molecular Biology
  • Microbiology
  • Structural Biology

Background:

  • Nascent polypeptide chains in ribosomes can regulate gene expression by inducing ribosome stalling.
  • The Staphylococcus aureus ErmCL leader peptide stalls translation in response to macrolide antibiotics like erythromycin, activating downstream macrolide resistance genes.
  • Understanding this mechanism is crucial for developing new antibiotic strategies.

Purpose of the Study:

  • To elucidate the structural basis of erythromycin-dependent ribosome stalling induced by the ErmCL leader peptide.
  • To reveal how the ErmCL nascent chain interacts with erythromycin and the ribosome to inhibit translation.

Main Methods:

  • Cryo-electron microscopy (cryo-EM) was used to determine the structure of the stalled ribosome.
  • High-resolution structural analysis (3.9 Å) was performed.

Main Results:

  • The cryo-EM structure reveals direct sensing of erythromycin by the ErmCL nascent chain within the ribosomal tunnel.
  • The ErmCL-drug interaction induces allosteric conformational changes in the peptidyltransferase center (PTC).
  • These PTC perturbations prevent aminoacyl-tRNA binding and inhibit peptide bond formation, causing translation arrest.

Conclusions:

  • The ErmCL leader peptide acts as a direct sensor for erythromycin, linking antibiotic presence to translational control.
  • The observed allosteric mechanism provides a molecular explanation for macrolide-induced ribosome stalling and resistance gene induction.
  • This study offers insights into bacterial antibiotic resistance and potential targets for novel therapeutics.

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