A Genetic Tool to Quantify trans-Translation Activity in Vivo - Université de Rennes Access content directly
Journal Articles Journal of Molecular Biology Year : 2017

A Genetic Tool to Quantify trans-Translation Activity in Vivo

Abstract

In bacteria, trans-translation is the main quality control mechanism for rescuing ribosomes arrested during translation. This key process is universally conserved and plays a critical role in the viability and virulence of many pathogens. We developed a reliable in vivo double-fluorescence reporter system for the simultaneous quantification of both trans-translation and the associated proteolysis activities in bacteria. The assay was validated using mutant bacteria lacking tmRNA, SmpB, and the ClpP protease. Both antisense tmRNA-binding RNA and a peptide mimicking the SmpB C-terminal tail proved to be potent inhibitors of trans-translation in vivo. The double-fluorescent reporter was also tested with KKL-35, an oxadiazole derivative that is supposed to be a promising trans-translation inhibitor, and it surprisingly turns out that trans-translation is not the only target of KKL-35 in vivo.
No file

Dates and versions

hal-01659526 , version 1 (08-12-2017)

Identifiers

Cite

Carlos Blanco, Reynald Gillet, Kevin Macé, Fanny Demay, Charlotte Guyomar, et al.. A Genetic Tool to Quantify trans-Translation Activity in Vivo. Journal of Molecular Biology, 2017, 429 (23), pp.3617-3625. ⟨10.1016/j.jmb.2017.10.007⟩. ⟨hal-01659526⟩
61 View
0 Download

Altmetric

Share

Gmail Facebook X LinkedIn More