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

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

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

In Escherichia coli, elongation factor G (EF-G), a key protein in translational elongation, is particularly susceptible to oxidation. We demonstrated previously that EF-G is inactivated upon formation of an intramolecular disulphide bond. However, the details of the mechanism by which the oxidation of EF-G inhibits the function of EF-G on the ribosome remain to be elucidated. When we oxidized EF-G with hydrogen peroxide, neither the insertion of EF-G into the ribosome nor single-cycle translocation activity in vitro was affected. However, the GTPase activity and the dissociation of EF-G from the ribosome were suppressed when EF-G was oxidized. The synthesis of longer peptides was suppressed to a greater extent than that of a shorter peptide when EF-G was oxidized. Thus, the formation of the disulphide bond in EF-G might interfere with the hydrolysis of GTP that is coupled with dissociation of EF-G from the ribosome and might thereby retard the turnover of EF-G within the translational machinery. When we added thioredoxin to the suppressed translation system that included oxidized EF-G, translational activity was almost immediately restored. We propose that oxidation of EF-G might provide a regulatory mechanism for transient and reversible suppression of translation in E. coli under oxidative stress.

Original languageEnglish
Pages (from-to)165-172
Number of pages8
JournalJournal of biochemistry
Volume158
Issue number2
DOIs
Publication statusPublished - 2014 Dec 26
Externally publishedYes

Keywords

  • EF-G
  • GTP hydrolysis
  • Oxidative stress
  • Protein synthesis
  • Redox regulation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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  • Cite this

    Nagano, T., Yutthanasirikul, R., Hihara, Y., Hisabori, T., Kanamori, T., Takeuchi, N., Ueda, T., & Nishiyama, Y. (2014). Oxidation of translation factor EF-G transiently retards the translational elongation cycle in Escherichia coli. Journal of biochemistry, 158(2), 165-172. https://doi.org/10.1093/jb/mvv026