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Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses
11:19

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

Published on: February 25, 2011

Role of antibiotic ligand in nascent peptide-dependent ribosome stalling

Nora Vázquez-Laslop1, Dorota Klepacki, Debbie C Mulhearn

  • 1Center for Pharmaceutical Biotechnology, University of Illinois, 900 South Ashland Avenue, Chicago, IL 60607, USA. nvazquez@uic.edu

Proceedings of the National Academy of Sciences of the United States of America
|June 15, 2011
PubMed

Insights

Cofactors, like antibiotics, are recognized with nascent peptides to arrest translation. This discovery reveals a new mechanism for regulating gene expression through precise molecular interactions within the ribosome.

Area of Science:

  • Molecular Biology
  • Bacterial Genetics
  • Biochemistry

Background:

  • Nascent peptides within the ribosome exit tunnel can induce translation arrest, a mechanism regulating gene expression in bacteria and eukaryotes.
  • This ribosome stalling is often modulated by small-molecule cofactors, but their precise role remains unclear.

Purpose of the Study:

  • To investigate the function of cofactors in programmed translation arrest.
  • To elucidate the mechanism by which antibiotics modulate ribosome stalling.

Main Methods:

  • Analysis of nascent peptide- and antibiotic-dependent ribosome stalling in bacteria controlling antibiotic resistance gene expression.
  • Investigating the impact of structural alterations in antibiotics on ribosome stalling efficiency.
  • Site-directed mutagenesis of 23S rRNA (C2610) to assess its role in sensing antibiotics.

Main Results:

  • Antibiotics are directly recognized as part of the translation-modulating signal, not just as external cues.
  • Minor structural changes in antibiotics significantly affect their ability to facilitate ribosome stalling, highlighting the importance of precise molecular interactions.
  • Mutation of 23S rRNA residue C2610 impairs nascent peptide- and antibiotic-dependent ribosome stalling.

Conclusions:

  • A new paradigm for programmed translation arrest is proposed, where small molecules (cofactors) are recognized alongside specific nascent peptides as a composite structure to induce arrest.
  • This mechanism suggests the ribosome can sense various cellular metabolites through similar composite recognition processes.

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