title: Evidence That Antibiotics Bind to Human Mitochondrial Ribosomal RNA Has Implications for Aminoglycoside Toxicity.
abstract: Aminoglycosides are a well known antibiotic family used to treat bacterial infections in humans and animals, but which can be toxic. By binding to the decoding site of helix44 of the small subunit RNA of the bacterial ribosome, the aminoglycoside antibiotics inhibit protein synthesis, cause misreading, or obstruct peptidyl-tRNA translocation. Although aminoglycosides bind helix69 of the bacterial large subunit RNA as well, little is known about their interaction with the homologous human helix69. To probe the role this binding event plays in toxicity, changes to thermal stability, base stacking, and conformation upon aminoglycoside binding to the human cytoplasmic helix69 were compared with those of the human mitochondrial and Escherichia coli helix69. Surprisingly, binding of gentamicin and kanamycin A to the chemically synthesized terminal hairpins of the human cytoplasmic, human mitochondrial, and E. coli helix69 revealed similar dissociation constants (1.3-1.7 and 4.0-5.4 μM, respectively). In addition, aminoglycoside binding enhanced conformational stability of the human mitochondrial helix69 by increasing base stacking. Proton one-dimensional and two-dimensional NMR suggested significant and specific conformational changes of human mitochondrial and E. coli helix69 upon aminoglycoside binding, as compared with human cytoplasmic helix69. The conformational changes and similar aminoglycoside binding affinities observed for human mitochondrial helix69 and E. coli helix69, as well as the increase in structural stability shown for the former, suggest that this binding event is important to understanding aminoglycoside toxicity.
authors.affiliation: From The RNA Institute and the Department of Biological Sciences, University at Albany, Albany, New York 12222.
authors.forename: Seoyeon
authors.initials: S
authors.lastname: Hong
authors.affiliation: From The RNA Institute and the Department of Biological Sciences, University at Albany, Albany, New York 12222.
authors.forename: Kimberly A
authors.initials: KA
authors.lastname: Harris
authors.affiliation: From The RNA Institute and the Department of Biological Sciences, University at Albany, Albany, New York 12222.
authors.forename: Kathryn D
authors.initials: KD
authors.lastname: Fanning
authors.affiliation: From The RNA Institute and the Department of Biological Sciences, University at Albany, Albany, New York 12222.
authors.forename: Kathryn L
authors.initials: KL
authors.lastname: Sarachan
authors.affiliation: From The RNA Institute and the Department of Biological Sciences, University at Albany, Albany, New York 12222.
authors.forename: Kyla M
authors.initials: KM
authors.lastname: Frohlich
authors.affiliation: From The RNA Institute and the Department of Biological Sciences, University at Albany, Albany, New York 12222 pagris@albany.edu.
authors.forename: Paul F
authors.initials: PF
authors.lastname: Agris
mesh_terms: D000900:Anti-Bacterial Agents / Q000737:chemistry*; D004926:Escherichia coli; D005839:Gentamicins / Q000737:chemistry*; D006801:Humans; D055029:Inverted Repeat Sequences; D007612:Kanamycin / Q000737:chemistry*; D012313:RNA / Q000737:chemistry*; D020871:RNA Stability; D012329:RNA, Bacterial / Q000737:chemistry; D000077278:RNA, Mitochondrial; D012335:RNA, Ribosomal / Q000737:chemistry*
chemical_list: D000900:Anti-Bacterial Agents; D005839:Gentamicins; D012329:RNA, Bacterial; D000077278:RNA, Mitochondrial; D012335:RNA, Ribosomal; D007612:Kanamycin; D012313:RNA
keywords: antibiotic action; circular dichroism (CD); nuclear magnetic resonance (NMR); ribosomal ribonucleic acid (rRNA) (ribosomal RNA); translation