Contribution of the second OB fold of ribosomal protein S1 from Escherichia coli to the recognition of TmRNA

Takahiro Okada, Iwona K. Wower, Jacek Wower, Christian W. Zwieb, Makoto Kimura

Research output: Contribution to journalArticle

15 Scopus citations

Abstract

Escherichia coli ribosomal protein S1 is composed of six repeating homologous oligonucleotide/oligosaccharide-binding fold (OB folds). In trans-translation, S1 plays a role in delivering transfer-messenger RNA (tmRNA) to stalled ribosomes. The second OB fold of S1 was found to be protected from tryptic digestion in the presence of tmRNA. Truncated S1 mutant Δ2, in which the first and second OB folds were deleted, showed significantly decreased tmRNA-binding activity. Furthermore, the E. coli S1 homolog (BS1) from Bacillus subtilis, which corresponds to the four C-terminal OB folds of E. coli S1, showed no interaction with E. coli tmRNA, as judged by the results of a gel shift assay. Surface plasmon resonance analysis revealed that mutant Δ2 and BS1 had decreased association rate constants (ka, 0.59 × 10 3M-1·S-1; and ka, 1.89 × 10 3M-1·S-1), while they retained the respective dissociation rate constants (kd, 0.67 × 10-3 S -1; and kd, 0.53 × 10-3 S-1), in comparison with wild-type protein S1 (ka, 3.32 × 103 M -1·S-1; and kd, 0.56 × 10-3 S -1). These results suggest that the second OB fold in protein S1 is essential for the recognition of tmRNA, while the four C-terminal OB folds play a role in stabilizing the S1-tmRNA complex.

Original languageEnglish (US)
Pages (from-to)2319-2325
Number of pages7
JournalBioscience, Biotechnology and Biochemistry
Volume68
Issue number11
DOIs
StatePublished - Nov 2004
Externally publishedYes

Keywords

  • Gel shift assay
  • Oligonucleotide/oligosaccharide-binding fold (OB fold)
  • Ribosomal protein S1
  • Trans-translation
  • Transfer-messenger RNA (tmRNA)

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

Fingerprint Dive into the research topics of 'Contribution of the second OB fold of ribosomal protein S1 from Escherichia coli to the recognition of TmRNA'. Together they form a unique fingerprint.

  • Cite this