ArfA recruits release factor 2 to rescue stalled ribosomes by peptidyl-tRNA hydrolysis in Escherichia coli

Yuhei Chadani, Koreaki Ito, Kazuhiro Kutsukake, Tatsuhiko Abo

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

The ribosomes stalled at the end of non-stop mRNAs must be rescued for productive cycles of cellular protein synthesis. Escherichia coli possesses at least three independent mechanisms that resolve non-productive translation complexes (NTCs). While tmRNA (SsrA) mediates trans-translation to terminate translation, ArfA (YhdL) and ArfB (YaeJ) induce hydrolysis of ribosome-tethered peptidyl-tRNAs. ArfB is a paralogue of the release factors (RFs) and directly catalyses the peptidyl-tRNA hydrolysis within NTCs. In contrast, the mechanism of the ArfA action had remained obscure beyond its ability to bind to the ribosome. Here, we characterized the ArfA pathway of NTC resolution in vitro and identified RF2 as a factor that cooperates with ArfA to hydrolyse peptidyl-tRNAs located in the P-site of the stalled ribosome. This reaction required the GGQ (Gly-Gly-Gln) hydrolysis motif, but not the SPF (Ser-Pro-Phe) codon-recognition sequence, of RF2 and was stimulated by tRNAs. From these results we suggest that ArfA binds to the vacant A-site of the stalled ribosome with possible aid from association with a tRNA, and then recruits RF2, which hydrolyses peptidyl-tRNA in a GGQ motif-dependent but codon-independent manner. In support of this model, the ArfA-RF2 pathway did not act on the SecM-arrested ribosome, which contains an aminoacyl-tRNA in the A-site.

Original languageEnglish
Pages (from-to)37-50
Number of pages14
JournalMolecular Microbiology
Volume86
Issue number1
DOIs
Publication statusPublished - Oct 2012

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Ribosomes
Hydrolysis
Escherichia coli
Transfer RNA
Codon
peptidyl-tRNA
Messenger RNA
Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

Cite this

ArfA recruits release factor 2 to rescue stalled ribosomes by peptidyl-tRNA hydrolysis in Escherichia coli. / Chadani, Yuhei; Ito, Koreaki; Kutsukake, Kazuhiro; Abo, Tatsuhiko.

In: Molecular Microbiology, Vol. 86, No. 1, 10.2012, p. 37-50.

Research output: Contribution to journalArticle

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