Oxidation of translation factor EF-G transiently retards the translational elongation cycle in<i>Escherichia coli</i>

Nagano, Takanori; Yutthanasirikul, Rayakorn; Hihara, Yukako; Hisabori, Toru; Kanamori, Takashi; Takeuchi, Naonobu; Ueda, Takuya; Nishiyama, Yoshitaka

doi:10.1093/jb/mvv026

Cited by 11 publications

(8 citation statements)

References 21 publications

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“…The authors further showed that in an E. coli in vitro translation system the use of the oxidized EF-Tu resulted in strongly inhibited translation. Similar findings were described for EF-G in both Synechocystis and E. coli ( Kojima et al, 2009 ; Nagano et al, 2015 ). EF-G catalyzes the tRNA/mRNA translocation step of elongation, where after a new peptide bond is formed between two amino acids, the ribosome moves along the mRNA to the next codon.…”

Section: The Transcribed Mrna Is Translated To Proteinssupporting

confidence: 85%

Oxidative Stress in Bacteria and the Central Dogma of Molecular Biology

Fasnacht

Polacek

2021

Front. Mol. Biosci.

140

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Ever since the “great oxidation event,” Earth’s cellular life forms had to cope with the danger of reactive oxygen species (ROS) affecting the integrity of biomolecules and hampering cellular metabolism circuits. Consequently, increasing ROS levels in the biosphere represented growing stress levels and thus shaped the evolution of species. Whether the ROS were produced endogenously or exogenously, different systems evolved to remove the ROS and repair the damage they inflicted. If ROS outweigh the cell’s capacity to remove the threat, we speak of oxidative stress. The injuries through oxidative stress in cells are diverse. This article reviews the damage oxidative stress imposes on the different steps of the central dogma of molecular biology in bacteria, focusing in particular on the RNA machines involved in transcription and translation.

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Section: The Transcribed Mrna Is Translated To Proteinssupporting

confidence: 85%

Oxidative Stress in Bacteria and the Central Dogma of Molecular Biology

Fasnacht

Polacek

2021

Front. Mol. Biosci.

140

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“…However, complex machinery can malfunction when its critical parts are damaged. Studies using model organisms from every domain of life have uncovered ROS-induced chemical alterations in aminoacyl-tRNA-synthetases and protein factors involved in the initiation, elongation, and termination of translation [8,14,15,16,17,18,19]. Exposure to oxidants affects posttranslational modifications and the stability of tRNAs [20,21,22].…”

Section: Introductionmentioning

confidence: 99%

The Impact of Oxidative Stress on Ribosomes: From Injury to Regulation

Shcherbik

Pestov

2019

Cells

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The ribosome is a complex ribonucleoprotein-based molecular machine that orchestrates protein synthesis in the cell. Both ribosomal RNA and ribosomal proteins can be chemically modified by reactive oxygen species, which may alter the ribosome′s functions or cause a complete loss of functionality. The oxidative damage that ribosomes accumulate during their lifespan in a cell may lead to reduced or faulty translation and contribute to various pathologies. However, remarkably little is known about the biological consequences of oxidative damage to the ribosome. Here, we provide a concise summary of the known types of changes induced by reactive oxygen species in rRNA and ribosomal proteins and discuss the existing experimental evidence of how these modifications may affect ribosome dynamics and function. We emphasize the special role that redox-active transition metals, such as iron, play in ribosome homeostasis and stability. We also discuss the hypothesis that redox-mediated ribosome modifications may contribute to adaptive cellular responses to stress.

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“…EF-Tu of Synechocystis includes a single cysteine residue, namely, Cys-82, and this residue corresponds to the residue that is strongly conserved. In EF-G of Synechocystis and E. coli, a specific pair of cysteine residues that is also conserved in various organisms is the target of oxidation by ROS (8,9,17,18). These observations together led us to postulate that Cys-82 in EF-Tu might be a potential target of oxidation by ROS.…”

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confidence: 99%

Oxidation of a Cysteine Residue in Elongation Factor EF-Tu Reversibly Inhibits Translation in the Cyanobacterium Synechocystis sp. PCC 6803

Yutthanasirikul¹,

Nagano²,

Jimbo³

et al. 2016

Journal of Biological Chemistry

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Translational elongation is susceptible to inactivation by reactive oxygen species (ROS) in the cyanobacterium Synechocystis sp. PCC 6803, and elongation factor G has been identified as a target of oxidation by ROS. In the present study we examined the sensitivity to oxidation by ROS of another elongation factor, EF-Tu. The structure of EF-Tu changes dramatically depending on the bound nucleotide. Therefore, we investigated the sensitivity to oxidation in vitro of GTP-and GDP-bound EF-Tu as well as that of nucleotide-free EF-Tu. Assays of translational activity with a reconstituted translation system from Escherichia coli revealed that GTP-bound and nucleotide-free EF-Tu were sensitive to oxidation by H 2 O 2 , whereas GDPbound EF-Tu was resistant to H 2 O 2 . The inactivation of EF-Tu was the result of oxidation of Cys-82, a single cysteine residue, and subsequent formation of both an intermolecular disulfide bond and sulfenic acid. Replacement of Cys-82 with serine rendered EF-Tu resistant to inactivation by H 2 O 2 , confirming that Cys-82 was a target of oxidation. Furthermore, oxidized EF-Tu was reduced and reactivated by thioredoxin. Gel-filtration chromatography revealed that some of the oxidized nucleotide-free EF-Tu formed large complexes of >30 molecules. Atomic force microscopy revealed that such large complexes dissociated into several smaller aggregates upon the addition of dithiothreitol. Immunological analysis of the redox state of EF-Tu in vivo showed that levels of oxidized EF-Tu increased under strong light. Thus, resembling elongation factor G, EF-Tu appears to be sensitive to ROS via oxidation of a cysteine residue, and its inactivation might be reversed in a redox-dependent manner.

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Oxidation of translation factor EF-G transiently retards the translational elongation cycle inEscherichia coli

Cited by 11 publications

References 21 publications

Oxidative Stress in Bacteria and the Central Dogma of Molecular Biology

Oxidative Stress in Bacteria and the Central Dogma of Molecular Biology

The Impact of Oxidative Stress on Ribosomes: From Injury to Regulation

Oxidation of a Cysteine Residue in Elongation Factor EF-Tu Reversibly Inhibits Translation in the Cyanobacterium Synechocystis sp. PCC 6803

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