The Interaction of Fusidic Acid with Peptidyl‐Transfer‐Ribonucleic‐Acid · Ribosome Complexes

Millan, María José San; Vázquez, David; Modoleli, Juan

doi:10.1111/j.1432-1033.1975.tb02318.x

Cited by 16 publications

(6 citation statements)

References 37 publications

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“…Puromycin reacts with peptidyl‐tRNA only after translocation of the peptidyl‐tRNA and tetracycline inhibits aminoacyl‐tRNA binding to ribosomes [32]. Viomycin inhibits translocation, but not GTP hydrolysis [33], and fusidic acid prevents the release of EF‐G–GDP from ribosomes but does not inhibit either translocation or GTP hydrolysis [34]. The data shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

Depletion of Escherichia coli 4.5S RNA leads to an increase in the amount of protein elongation factor EF‐G associated with ribosomes

Nakamura

Fujii

Shibata

et al. 1999

European Journal of Biochemistry

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In Escherichia coli, 4.5S RNA is found in complexes with both protein translocation protein, Ffh (a bacterial homolog of mammalian SRP54) and protein synthesis elongation factor G (EF‐G). To analyze the function of 4.5S RNA in translation, we initially assessed the sensitivity of the association of 4.5S RNA with the ribosome after treatment with antibiotics that affect various stages of protein synthesis. Fusidic acid and viomycin caused 4.5S RNA to cosediment with the 70S ribosomal fraction, indicating that 4.5S RNA enters the ribosome before ribosomal translocation and release of EF‐G–GDP from the ribosome. On the other hand, depletion of 4.5S RNA led to the retention of a significant amount of EF‐G on 70S ribosomes. In addition, 4.5S RNA shares a conserved decanucleotide sequence (58GAAGCAGCCA67) motif with the characterized EF‐G‐binding site at positions 1068–1077 on 23S RNA. We therefore examined by gel mobility‐shift assay whether or not mutations in the domain‐IV region of 4.5S RNA, including this conserved motif, disturb the binding of EF‐G to 23S RNA. Any mutation at the C62, G64 or A67 residues within this motif abolished competition activity. Therefore, we propose that 4.5S RNA is concerned with the mode of association of EF‐G with the ribosomes. Moreover, this function depends on the secondary structure of 4.5S RNA as well as a ten‐base sequence conserved between the two RNAs.

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Section: Resultsmentioning

confidence: 99%

Depletion of Escherichia coli 4.5S RNA leads to an increase in the amount of protein elongation factor EF‐G associated with ribosomes

Nakamura

Fujii

Shibata

et al. 1999

European Journal of Biochemistry

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“…3) could be construed as evidence for two binding sites for the antibiotic with different affinities. Alternatively, since translocation occurs on ribosomes with bound peptidyl-tRNA, but the fusidic acid-stabilized complex forms mainly on free ribosomes [14,31], the different sensitivities could be due to a single viomycin-binding site whose affinity for the antibiotic would be increased in the presence of peptidyl-tRNA. Evidently, binding studies with radioactively labelled viomycin would clarify these points and help elucidate whether or not the dual action of the antibiotic on initiation and elongation …”

Section: Discussionmentioning

confidence: 99%

The Inhibition of Ribosomal Translocation by Viomycin

Modolfll¹,

Vázquez²

1977

European Journal of Biochemistry

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119

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The basic peptide antibiotic viomycin rapidly inhibits polypeptide chain elongation when added to purified endogenous Escherichia coli polysomes actively engaged in polypeptide synthesis. After inhibition is established, the polysomal nascent peptide chains react only slowly with puromycin, while the chains of uninhibited polysomes or tetracycline-blocked polysomes react very rapidly. Since viomycin does not inhibit the peptidyl transferase reaction, these results indicate that the antibiotic confines peptidyl-tRNA to the ribosomal acceptor site and, consequently, that it blocks polypeptide chain elongation by inhibiting ribosomal translocation.Viomycin at concentrations strongly inhibitory for translocation (1 0 pM) does not impair the elongation-factor (EF)-G-plus-ribosome-dependent GTP hydrolysis, the formation of GDP . EF-G . ribosome . fusidic-acid complex and guanyl-5'-yl-methylene-diphosphonate . EF-G . ribosome complex, or the non-enzymic binding of AcPhe-tRNA to the ribosomal donor site. In contrast, viomycin strongly inhibits the non-enzymic translocation and the release of AcPhe-tRNA from the ribosomal acceptor site promoted by depletion of NHZ ions. It is suggested that viomycin inhibits the movement of peptidyl-tRNA associated with translocation by interfering with its exit from the ribosomal acceptor site. Viomycin at high concentrations (10 -I00 pM) partially inhibits the EF-G-plus-ribosome-dependent GTP hydrolysis and the turnover of the GDP . EF-G ribosome . fusidic-acid complex. These effects, however, are probably unrelated to the inhibition of ribosomal translocation.

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“…The uncoiling was also induced with 15 pg fucidin/ml growth medium. RICHTER et al 1971;MAZUMDER 1975;MILLAN et al 1975). When added to lymphoid cell in G, phase fucidin (the sodium salt of fusidic acid) produces banded or uncoiled chromosomes in the treated cells.…”

Section: Resultsmentioning

confidence: 99%