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Proc Natl Acad Sci U S A. 2017 Dec 19;114(51):E10899-E10908. doi: 10.1073/pnas.1715501114. Epub 2017 Dec 5.

Aminoglycoside interactions and impacts on the eukaryotic ribosome.

Author information

1
Department of Integrated Structural Biology, Institut de Génétique et de Biologie Moléculaire et Cellulaire, INSERM U964, CNRS UMR7104, Université de Strasbourg, 67404 Illkirch, France.
2
Department of Physiology and Biophysics, Weill Cornell Medicine, New York, NY 10065.
3
Department of Chemistry and Biochemistry, Utah State University, Logan, UT 84322.
4
Département de Physique, Faculté des Sciences et des Technologies, Université de Lorraine, 54506 Vandoeuvre-lès-Nancy, France.
5
Tri-Institutional PhD Training Program in Chemical Biology, Weill Cornell Medicine, Rockefeller University, Memorial Sloan-Kettering Cancer Center, New York, NY 10065.
6
Department of Integrated Structural Biology, Institut de Génétique et de Biologie Moléculaire et Cellulaire, INSERM U964, CNRS UMR7104, Université de Strasbourg, 67404 Illkirch, France; marat@igbmc.fr scb2005@med.cornell.edu gula@igbmc.fr.
7
Department of Physiology and Biophysics, Weill Cornell Medicine, New York, NY 10065; marat@igbmc.fr scb2005@med.cornell.edu gula@igbmc.fr.

Abstract

Aminoglycosides are chemically diverse, broad-spectrum antibiotics that target functional centers within the bacterial ribosome to impact all four principle stages (initiation, elongation, termination, and recycling) of the translation mechanism. The propensity of aminoglycosides to induce miscoding errors that suppress the termination of protein synthesis supports their potential as therapeutic interventions in human diseases associated with premature termination codons (PTCs). However, the sites of interaction of aminoglycosides with the eukaryotic ribosome and their modes of action in eukaryotic translation remain largely unexplored. Here, we use the combination of X-ray crystallography and single-molecule FRET analysis to reveal the interactions of distinct classes of aminoglycosides with the 80S eukaryotic ribosome. Crystal structures of the 80S ribosome in complex with paromomycin, geneticin (G418), gentamicin, and TC007, solved at 3.3- to 3.7-Å resolution, reveal multiple aminoglycoside-binding sites within the large and small subunits, wherein the 6'-hydroxyl substituent in ring I serves as a key determinant of binding to the canonical eukaryotic ribosomal decoding center. Multivalent binding interactions with the human ribosome are also evidenced through their capacity to affect large-scale conformational dynamics within the pretranslocation complex that contribute to multiple aspects of the translation mechanism. The distinct impacts of the aminoglycosides examined suggest that their chemical composition and distinct modes of interaction with the ribosome influence PTC read-through efficiency. These findings provide structural and functional insights into aminoglycoside-induced impacts on the eukaryotic ribosome and implicate pleiotropic mechanisms of action beyond decoding.

KEYWORDS:

PTC read-through; aminoglycosides; protein synthesis; ribosome; translation

PMID:
29208708
PMCID:
PMC5754804
DOI:
10.1073/pnas.1715501114
[Indexed for MEDLINE]
Free PMC Article

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