FEBS Letters
Volume 514, Issue 1 , Pages 70-73, 6 March 2002

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

Edited by Lev Kisselev

  • Olga N Avdeeva

      Affiliations

    • Department of Chemistry, Moscow State University, Moscow 119899, Russia
    • ,
  • Alexander G Myasnikov

      Affiliations

    • Department of Chemistry, Moscow State University, Moscow 119899, Russia
    • ,
  • Petr V Sergiev

      Affiliations

    • Department of Chemistry, Moscow State University, Moscow 119899, Russia
    • Corresponding Author InformationCorresponding author. Fax: (7)-095-9393181
    • ,
  • Alexey A Bogdanov

      Affiliations

    • Department of Chemistry, Moscow State University, Moscow 119899, Russia
    • ,
  • Richard Brimacombe

      Affiliations

    • Max-Planck Institut fur Molekulare Genetik AG-Ribosomen, Ihnestrasse 73, 14195 Berlin, Germany
    • ,
  • Olga A Dontsova

      Affiliations

    • Department of Chemistry, Moscow State University, Moscow 119899, Russia

Received 20 October 2001; received in revised form 24 October 2001; accepted 28 October 2001.

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.

Keywords:  Signal recognition particle, Ribosome, Translation arrest, Ffh, 4.5S RNA

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PII: S0014-5793(02)02332-3

FEBS Letters
Volume 514, Issue 1 , Pages 70-73, 6 March 2002

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