Formation of Fusidic Acid-G Factor-GDP-Ribosome Complex and the Relationship to the Inhibition of GTP Hydrolysis

Okura, Akira; Kinoshita, Tadatoshi; Tanaka, Nobuo

doi:10.7164/antibiotics.24.655

Cited by 33 publications

(19 citation statements)

References 23 publications

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“…13AB). On the other hand, FA, which prevents the rapid turnover of elongation factor G (EF-G) from the ribosome via binding to the EF-G-ribosome complex 26,27 , affects the ribosome movement more reversibly 28 . In this case, we found ER to decrease gradually with sub-lethal doses of FA treatment (Supplementary Fig.…”

Section: Results and Analysismentioning

confidence: 99%

Reduction of translating ribosomes enables Escherichia coli to maintain elongation rates during slow growth

et al. 2016

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Bacteria growing in different conditions experience a broad range of demand on the rate of protein synthesis which profoundly affects cellular resource allocation. During fast growth, protein synthesis is long known to be modulated by adjusting the ribosome content, with the vast majority of ribosomes engaged at a near-maximal rate of elongation. Here we characterized protein synthesis by E. coli systematically, focusing on slow growth conditions. We establish that the translational elongation rate decreases as growth slows down, exhibiting a Michaelis-Menten dependence on the abundance of the cellular translational apparatus. However, an appreciable elongation rate is maintained even towards zero growth including the stationary phase. This maintenance, critical for timely protein synthesis in harsh environments, is accompanied by a drastic reduction in the fraction of active ribosomes. Interestingly, well-known antibiotics such as chloramphenicol also cause substantial reduction in the pool of active ribosomes, instead of slowing down translational elongation as commonly thought.

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Section: Results and Analysismentioning

confidence: 99%

Reduction of translating ribosomes enables Escherichia coli to maintain elongation rates during slow growth

et al. 2016

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“…It was known that EF-G formed a stable complex with the ribosome in the presence of GTP (or GDP) and fusidic acid (65)(66)(67)(68)(69). As shown in Fig.…”

Section: Elongationmentioning

confidence: 97%

Chemistry and biology ofE. coli ribosomal protein L12

Brot

Weissbach

1981

Mol Cell Biochem

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E. coli ribosomal protein L12, because of its unique features, has been studied in more detail than perhaps any of the other ribosomal proteins. Unlike the other ribosomal proteins that are generally present in stoichiometric amounts, there are four copies of L12 per ribosome, some of which are acetylated on the N-terminal serine. The acetylated species, referred to as L7, has not been shown, as yet, to possess any different biological activity than L12. A specific enzyme that acetylates L12 to form L7, using acetyl-CoA as the acetyl donor, has been purified from E. coli extracts. L12 is also unique in that it does not contain cysteine, tryptophan, histidine, or tyrosine, is very acidic (pI: 4.85) and has a high content of ordered secondary structure (approximately 50%). The protein is normally found in solution as a dimer and also forms a tight complex with ribosomal protein L10. There are three methionine residues in L12, located in the N-terminal region of the protein, one or more of which are essential for biological activity. Oxidation of the methionines to methionine sulfoxide prevents dimer formation and inactivates the protein. The four copies of L12 are located in the crest region(s) of the 50S ribosomal subunit. There is good evidence that the soluble factors, such as IF-2, EF-Tu, EF-G and RF, interact with L12 on the ribosome during the process of protein synthesis. This interaction is essential for the proper functioning of each of the factors and for GTP hydrolysis associated with the individual partial reactions of protein synthesis. The L12 gene is located on an operon that contains the genes for L10 and beta beta' subunits of RNA polymerase at about 88 min on the bacterial chromosome. DNA-directed in vitro systems have been used to study the unique regulation of the expression of these genes. Autogenous regulation, translational control, and transcription attenuation are regulatory mechanisms that function to control the synthesis of these proteins.

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“…Among these is the steroid-like antibiotic fusidic acid (FA), 3 which stalls the ribosome in complex with EF-G and GDP (3). FA has low affinity to EF-G off the ribosome but forms a strong complex with ribosome-bound EF-G (3,4). EF-G plays double roles in bacterial protein synthesis by both catalyzing mRNA translocation during the peptide elongation cycle and, aided by ribosome recycling factor, splitting the post-termination ribosome into subunits (5) for a new round of initiation of mRNA translation (6,7).…”

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confidence: 99%

Fusidic Acid Targets Elongation Factor G in Several Stages of Translocation on the Bacterial Ribosome

Borg

Holm

Shiroyama

et al. 2015

Journal of Biological Chemistry

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Background: FA inhibits ribosomal elongation and recycling. Results: The mechanism of FA inhibition of peptide elongation was determined. Conclusion: FA acts as a strong inhibitor by targeting several EF-G-containing states of the peptide elongation cycle. Significance: This study places FA-stalled ribosomal structures in a functional context and enables in vivo modeling of FA action. Techniques for studying slow inhibitors were developed.

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Formation of Fusidic Acid-G Factor-GDP-Ribosome Complex and the Relationship to the Inhibition of GTP Hydrolysis

Cited by 33 publications

References 23 publications

Reduction of translating ribosomes enables Escherichia coli to maintain elongation rates during slow growth

Reduction of translating ribosomes enables Escherichia coli to maintain elongation rates during slow growth

Chemistry and biology ofE. coli ribosomal protein L12

Fusidic Acid Targets Elongation Factor G in Several Stages of Translocation on the Bacterial Ribosome

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