The structural mechanism of the inhibition of archaeal RelE toxin by its cognate RelB antitoxin

Masaaki Shinohara, Jin Xu Guo, Misako Mori, Takashi Nakashima, Hisanori Takagi, Etsuko Nishimoto, Shoji Yamashita, Kouhei Tsumoto, Yoshimitsu Kakuta, Makoto Kimura

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The archaeal toxin, aRelE, in the hyperthermophilic archaeon Pyrococcus horikoshii OT3 inhibits protein synthesis, whereas its cognate antitoxin, aRelB, neutralizes aRelE activity by forming a non-toxic complex, aRelB-aRelE. The structural mechanism whereby aRelB neutralizes aRelE activity was examined by biochemical and biophysical analyses. Overexpression of aRelB with an aRelE mutant (ΔC6), in which the C-terminal residues critical for aRelE activity were deleted, in Escherichia coli allowed a stable complex, aRelB-ΔC6, to be purified. Isothermal titration of aRelE or ΔC6 with aRelB indicated that the association constant (Ka) of wild-type aRelB-aRelE is similar to that of aRelB-ΔC6, demonstrating that aRelB makes little contact with the C-terminal active site of aRelE. Overexpression of deletion mutants of aRelB with aRelE indicated that either the N-terminal (pos 1-27) or C-terminal (pos. 50-67) fragment of aRelB is sufficient to counteract the toxicity of aRelE in E. coli cells and the second α-helix (α2) in aRelB plays a critical role in forming a stable complex with aRelE. The present results demonstrate that aRelB, as expected from its X-ray structure, precludes aRelE from entering the ribosome, wrapping around the molecular surface of aRelE.

Original languageEnglish
Pages (from-to)346-351
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume400
Issue number3
DOIs
Publication statusPublished - Sep 1 2010

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Antitoxins
Escherichia coli
Pyrococcus horikoshii
Archaea
Titration
Ribosomes
Toxicity
Catalytic Domain
X-Rays
Association reactions
X rays
Proteins

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

The structural mechanism of the inhibition of archaeal RelE toxin by its cognate RelB antitoxin. / Shinohara, Masaaki; Guo, Jin Xu; Mori, Misako; Nakashima, Takashi; Takagi, Hisanori; Nishimoto, Etsuko; Yamashita, Shoji; Tsumoto, Kouhei; Kakuta, Yoshimitsu; Kimura, Makoto.

In: Biochemical and Biophysical Research Communications, Vol. 400, No. 3, 01.09.2010, p. 346-351.

Research output: Contribution to journalArticle

Shinohara, Masaaki ; Guo, Jin Xu ; Mori, Misako ; Nakashima, Takashi ; Takagi, Hisanori ; Nishimoto, Etsuko ; Yamashita, Shoji ; Tsumoto, Kouhei ; Kakuta, Yoshimitsu ; Kimura, Makoto. / The structural mechanism of the inhibition of archaeal RelE toxin by its cognate RelB antitoxin. In: Biochemical and Biophysical Research Communications. 2010 ; Vol. 400, No. 3. pp. 346-351.
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AU - Nishimoto, Etsuko

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