Related Experiment Video
Updated: Nov 5, 2025
Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses
Published on: February 25, 2011
Context-specific action of macrolide antibiotics on the eukaryotic ribosome
Maxim S Svetlov1,2, Timm O Koller3, Sezen Meydan1,4,5
1Center for Biomolecular Sciences, University of Illinois at Chicago, Chicago, IL, USA.
Abstract:
Macrolide antibiotics bind in the nascent peptide exit tunnel of the bacterial ribosome and prevent polymerization of specific amino acid sequences, selectively inhibiting translation of a subset of proteins. Because preventing translation of individual proteins could be beneficial for the treatment of human diseases, we asked whether macrolides, if bound to the eukaryotic ribosome, would retain their context- and protein-specific action. By introducing a single mutation in rRNA, we rendered yeast Saccharomyces cerevisiae cells sensitive to macrolides. Cryo-EM structural analysis showed that the macrolide telithromycin binds in the tunnel of the engineered eukaryotic ribosome. Genome-wide analysis of cellular translation and biochemical studies demonstrated that the drug inhibits eukaryotic translation by preferentially stalling ribosomes at distinct sequence motifs. Context-specific action markedly depends on the macrolide structure. Eliminating macrolide-arrest motifs from a protein renders its translation macrolide-tolerant. Our data illuminate the prospects of adapting macrolides for protein-selective translation inhibition in eukaryotic cells.
Insights
Macrolide antibiotics can be engineered to selectively inhibit protein translation in eukaryotic cells. This discovery opens new avenues for developing targeted therapies by controlling specific protein production.
Area of Science:
- Molecular Biology
- Biochemistry
- Genetics
Background:
- Macrolide antibiotics target bacterial ribosomes, inhibiting protein synthesis by binding to the nascent peptide exit tunnel.
- Selective translation inhibition of specific proteins holds therapeutic potential for human diseases.
Purpose of the Study:
- To investigate if macrolides can selectively inhibit protein translation in eukaryotic ribosomes.
- To explore the potential of macrolides as tools for protein-specific translation control in eukaryotes.
Main Methods:
- Engineered yeast Saccharomyces cerevisiae cells for macrolide sensitivity via an rRNA mutation.
- Utilized Cryo-electron microscopy (Cryo-EM) to visualize macrolide binding in eukaryotic ribosomes.
- Conducted genome-wide translation analysis and biochemical assays to assess drug effects.
Main Results:
- Telithromycin was observed to bind within the engineered eukaryotic ribosome's tunnel.
- Macrolides were found to stall eukaryotic translation at specific sequence motifs in a context-dependent manner.
- Protein translation became macrolide-tolerant when specific arrest motifs were removed.
Conclusions:
- Macrolides can be adapted to achieve protein-selective translation inhibition in eukaryotic cells.
- The structural context and macrolide chemical structure dictate the specificity of translation inhibition.
- This research highlights the potential of macrolides for targeted therapeutic applications in eukaryotes.
Related Concept Videos
01:23Types of RNA
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...
01:20Types of RNA
RNA Performs Diverse...
01:29Translational Regulation
02:53Ribosomal RNA Synthesis
Ribosome biogenesis begins with the synthesis of 5S and 45S pre-rRNAs by distinct RNA polymerases. The primary transcripts are extensively processed and modified before they are bound and folded by ribosomal proteins and assembly factors,...
02:53Ribosomal RNA Synthesis
01:23Stringent Response in E. coli