The translocation inhibitor tuberactinomycin binds to nucleic acids and blocks the in vitro assembly of 50S subunits

Yamada, Takeshi; Teshima, Tadashi; Shiba, Tetsuo; Nierhaus, Knud H.

doi:10.1093/nar/8.23.5767

Cited by 11 publications

(5 citation statements)

References 18 publications

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“…Here, two speculative conclusions can be made. First, the nearly complete inhibition of translocation observed at both high viomycin and neomycin concentrations, where previous data17,35,41 and our titration experiments argue multiple drug molecules may be bound, suggests that primary and secondary binding site occupancy may operate additively with respect to tRNA dynamics but synergistically on the extent of translocation inhibition. Second, the binding of antibiotics outside of the h44 decoding site, which have the net effect of stabilizing hybridstate tRNA configurations, may exhibit more pronounced translocation inhibition.…”

Section: Resultssupporting

confidence: 52%

Aminoglycoside activity observed on single pre-translocation ribosome complexes

et al. 2009

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Aminoglycoside-class antibiotics bind directly to ribosomal RNA, imparting pleiotropic effects on ribosome function. Despite in-depth structural investigations of aminoglycoside–RNA oligonucleotide and aminoglycoside-ribosome interactions, mechanisms explaining the unique ribosome inhibition profiles of chemically similar aminoglycosides remain elusive. Here, using single-molecule fluorescence resonance energy transfer (smFRET) methods, we show that high-affinity aminoglycoside binding to the conserved decoding site region of the functional pre-translocation ribosome complex specifically remodels the nature of intrinsic dynamic processes within the particle. The extents of these effects, which are distinct for each member of the aminoglycoside class, strongly correlate with their inhibition of EF-G–catalyzed translocation. Neomycin, a 4,5-linked amino-glycoside, binds with lower affinity to one or more secondary binding sites, mediating distinct structural and dynamic perturbations that further enhance translocation inhibition. These new insights help explain why closely related aminoglycosides elicit pleiotropic translation activities and demonstrate the potential utility of smFRET as a tool for dissecting the mechanisms of antibiotic action.

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Section: Resultssupporting

confidence: 52%

Aminoglycoside activity observed on single pre-translocation ribosome complexes

et al. 2009

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“…Here, two speculative conclusions can be made. First, the nearly complete inhibition of translocation observed at both high viomycin and neomycin concentrations, where previous data 17,35,41 and our titration experiments argue multiple drug molecules may be bound, suggests that primary and secondary binding site occupancy may operate additively with respect to tRNA dynamics but synergistically on the extent of translocation inhibition. Second, the binding of antibiotics outside of the h44 decoding site, which have the net effect of stabilizing hybrid-state tRNA configurations, may exhibit more pronounced translocation inhibition.…”

Section: Resultssupporting

confidence: 52%

Erratum: Aminoglycoside activity observed on single pre-translocation ribosome complexes

Feldman¹,

Terry²,

Altman³

et al. 2010

Nat Chem Biol

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Aminoglycoside-class antibiotics bind directly to ribosomal RNA, imparting pleiotropic effects on ribosome function. Despite in-depth structural investigations of aminoglycoside–RNA oligonucleotide and aminoglycoside-ribosome interactions, mechanisms explaining the unique ribosome inhibition profiles of chemically similar aminoglycosides remain elusive. Here, using single-molecule fluorescence resonance energy transfer (smFRET) methods, we show that high-affinity aminoglycoside binding to the conserved decoding site region of the functional pre-translocation ribosome complex specifically remodels the nature of intrinsic dynamic processes within the particle. The extents of these effects, which are distinct for each member of the aminoglycoside class, strongly correlate with their inhibition of EF-G–catalyzed translocation. Neomycin, a 4,5-linked aminoglycoside, binds with lower affinity to one or more secondary binding sites, mediating distinct structural and dynamic perturbations that further enhance translocation inhibition. These new insights help explain why closely related aminoglycosides elicit pleiotropic translation activities and demonstrate the potential utility of smFRET as a tool for dissecting the mechanisms of antibiotic action.

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“…The relatively low binding of tuberactinomycin to nucleic acids observed here is due to the higher ionic strength we applied in these equilibrium dialysis experiments in contrast to the previous data [12]. Increasing the ionic strength Maximal stimulation of the binding of tuberactinomycin to programmed ribosomes was achieved by the addition of a two-molar excess of tRNAPhe (375 pmol tRNAPhe, see table 1).…”

Section: Resultscontrasting

confidence: 53%

tRNA binding to programmed ribosomes increases the ribosomal affinity for tuberactinomycin O

et al. 1985

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The binding of i4C-labelled tuberactinomycin 0 was analysed in equilibrium dialysis cells. The ionic conditions and the concentration of the labelled drug used in the binding assays allowed the binding of just one drug molecule per non-programmed ribosome. Under these conditions, the occupation of the ribosomal P-site by deacylated tRNAP" in the presence. of poly(U) increased the amount of [%]tuberactinomycin 0 bound by a factor of two. Kanamycin, gentamicin and neomycin reduced the binding of tuberactinomycin 0, whereas chloramphenicol, tetracycline, streptomycin and puromycin had no effect. A stimulation of the binding of tuberactinomycin 0 was found upon addition of erythromycin.

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The translocation inhibitor tuberactinomycin binds to nucleic acids and blocks the in vitro assembly of 50S subunits

Cited by 11 publications

References 18 publications

Aminoglycoside activity observed on single pre-translocation ribosome complexes

Aminoglycoside activity observed on single pre-translocation ribosome complexes

Erratum: Aminoglycoside activity observed on single pre-translocation ribosome complexes

tRNA binding to programmed ribosomes increases the ribosomal affinity for tuberactinomycin O

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