Molecular Mechanism of Drug-Dependent Ribosome Stalling
- 1 April 2008
- journal article
- research article
- Published by Elsevier BV in Molecular Cell
- Vol. 30 (2), 190-202
- https://doi.org/10.1016/j.molcel.2008.02.026
Abstract
Inducible expression of the erm erythromycin resistance genes relies on drug-dependent ribosome stalling. The molecular mechanisms underlying stalling are unknown. We used a cell-free translation system to elucidate the contribution of the nascent peptide, the drug, and the ribosome toward formation of the stalled complex during translation of the ermC leader cistron. Toe-printing mapping, selective amino acid labeling, and mutational analyses revealed the peptidyl transferase center (PTC) as the focal point of the stalling mechanism. In the ribosome exit tunnel, the C-terminal sequence of the nascent peptide, critical for stalling, is in the immediate vicinity of the universally conserved A2062 of 23S rRNA. Mutations of this nucleotide eliminate stalling. Because A2062 is located in the tunnel, it may trigger a conformational change in the PTC, responding to the presence of a specific nascent peptide. The cladinose-containing macrolide antibiotic in the tunnel positions the nascent peptide for interaction with the tunnel sensory elements.Keywords
This publication has 55 references indexed in Scilit:
- Induction of erm (C) Expression by Noninducing AntibioticsAntimicrobial Agents and Chemotherapy, 2008
- Efficient protein selection based on ribosome display system with purified componentsBiochemical and Biophysical Research Communications, 2007
- Translation Arrest Requires Two-Way Communication between a Nascent Polypeptide and the RibosomeMolecular Cell, 2006
- Genetically Encoded but Nonpolypeptide Prolyl-tRNA Functions in the A Site for SecM-Mediated Ribosomal StallMolecular Cell, 2006
- Features of Ribosome-Peptidyl-tRNA Interactions Essential for Tryptophan Induction of tna Operon ExpressionMolecular Cell, 2005
- Instruction of Translating Ribosome by Nascent PeptideScience, 2002
- Binding Site of Macrolide Antibiotics on the Ribosome: New Resistance Mutation Identifies a Specific Interaction of Ketolides with rRNAJournal of Bacteriology, 2001
- The Structural Basis of Ribosome Activity in Peptide Bond SynthesisScience, 2000
- Defining the Structural Requirements for a Helix in 23 S Ribosomal RNA that Confers Erythromycin ResistanceJournal of Molecular Biology, 1989
- ermC leader peptide: Amino acid sequence critical for induction by translational attenuationJournal of Molecular Biology, 1989