Construction of the 'minimal' SRP that interacts with the translating ribosome but not with specific membrane receptors in Escherichia coli

Olga N. Avdeeva, Alexander G. Myasnikov, Petr V. Sergiev, Alexey A. Bogdanov, Richard Brimacombe, Olga A. Dontsova

Research output: Contribution to journalArticlepeer-review

16 Citations (Scopus)

Abstract

Escherichia coli signal recognition particle (SRP) consists of 4.5S RNA and Ffh protein. In contrast to eukaryotes, it remains unclear whether translation arrest takes place in prokaryotic cells. To study this problem we constructed a fusion of the M domain of Ffh protein with a cleavable affinity tag. This mutant Ffh, in a complex with 4.5S RNA, can bind signal peptide at the translating ribosome but is unable to bind the membrane. This SRP-ribosome complex should accumulate in the cell if translation is arrested. To test this, the complex was purified from the cells by ultracentrifugation and affinity chromatography. The composition of the complex was analyzed and found to consist of ribosomal RNAs and proteins, the Ffh M domain and 4.5S RNA. The accumulation of this complex in the cell in significant amounts indicated that SRP-mediated translation arrest did occur in bacterial cells.

Original languageEnglish
Pages (from-to)70-73
Number of pages4
JournalFEBS Letters
Volume514
Issue number1
DOIs
Publication statusPublished - 6 Mar 2002
Externally publishedYes

Keywords

  • 4.5S RNA
  • Ffh
  • Ribosome
  • Signal recognition particle
  • Translation arrest

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