The Termination Phase in Protein Synthesis is not Obligatorily Followed by the RRF/EF-G-Dependent Recycling Phase

Bo Qin, Hiroshi Yamamoto, Takuya Ueda, Umesh Varshney, Knud H. Nierhaus

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

It is general wisdom that termination of bacterial protein synthesis is obligatorily followed by recycling governed by the factors ribosomal recycling factor (RRF), EF-G, and IF3, where the ribosome dissociates into its subunits. In contrast, a recently described 70S-scanning mode of initiation holds that after termination, scanning of 70S can be triggered by fMet-tRNA to the initiation site of a downstream cistron. Here, we analyze the apparent conflict. We constructed a bicistronic mRNA coding for luciferases and showed with a highly resolved in vitro system that the expression of the second cistron did not at all depend on the presence of active RRF. An in vivo analysis cannot be performed in a straightforward way, since RRF is essential for viability and therefore, the RRF gene cannot be knocked out. However, we found an experimental window, where the RRF amount could be reduced to below 2.5%, and in this situation, the expression of the second cistron of a bicistronic luciferase mRNA was only moderately reduced. Both in vitro and in vivo results suggested that RRF-dependent recycling is not an obligatory step after termination, in agreement with the previous findings concerning 70S-scanning initiation. In this view, recycling after termination is a special case of the general RRF function, which happens whenever fMet-tRNA is not available for triggering 70S scanning.

Original languageEnglish
Pages (from-to)3577-3587
Number of pages11
JournalJournal of Molecular Biology
Volume428
Issue number18
DOIs
Publication statusPublished - 2016 Sep 11
Externally publishedYes

Fingerprint

Peptide Elongation Factor G
Recycling
Proteins
Luciferases
Genes
Messenger RNA
Bacterial Proteins
Ribosomes

Keywords

  • protein synthesis
  • recycling phase not obligatory
  • ribosome recycling
  • RRF function
  • termination of protein synthesis

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

The Termination Phase in Protein Synthesis is not Obligatorily Followed by the RRF/EF-G-Dependent Recycling Phase. / Qin, Bo; Yamamoto, Hiroshi; Ueda, Takuya; Varshney, Umesh; Nierhaus, Knud H.

In: Journal of Molecular Biology, Vol. 428, No. 18, 11.09.2016, p. 3577-3587.

Research output: Contribution to journalArticle

Qin, Bo ; Yamamoto, Hiroshi ; Ueda, Takuya ; Varshney, Umesh ; Nierhaus, Knud H. / The Termination Phase in Protein Synthesis is not Obligatorily Followed by the RRF/EF-G-Dependent Recycling Phase. In: Journal of Molecular Biology. 2016 ; Vol. 428, No. 18. pp. 3577-3587.
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