4.4 Article

Safe and easy in vitro evaluation of tmRNA-SmpB-mediated trans-translation from ESKAPE pathogenic bacteria

期刊

RNA
卷 27, 期 11, 页码 1390-1399

出版社

COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT
DOI: 10.1261/rna.078773.121

关键词

trans-translation; tmRNA; ESKAPE; antibiotics; HTS; ribosome

资金

  1. Agence Nationale pour la Recherche
  2. SATT Ouest-Valorisation [DV 2552, DV 3506]

向作者/读者索取更多资源

Trans-translation is a major quality control system in bacteria for rescuing stalled ribosomes and is a promising target for novel antibiotic development. Various in vitro and in vivo systems have been created for evaluating trans-translational activity, but the focus of current work is on safe and easy in vitro evaluation specifically for pathogenic bacteria.
In bacteria, trans-translation is the major quality control system for rescuing stalled ribosomes. It is mediated by tmRNA, a hybrid RNA with properties of both a tRNA and a mRNA, and the small protein SmpB. Because trans-translation is absent in eukaryotes but necessary for bacterial fitness or survival, it is a promising target for the development of novel antibiotics. To facilitate screening of chemical libraries, various reliable in vitro and in vivo systems have been created for assessing trans-translational activity. However, the aim of the current work was to permit the safe and easy in vitro evaluation of trans-translation from pathogenic bacteria, which are obviously the ones we should be targeting. Based on green fluorescent protein (GFP) reassembly during active trans-translation, we have created a cell-free assay adapted to the rapid evaluation of trans-translation in ESKAPE bacteria, with 24 different possible combinations. It can be used for easy high-throughput screening of chemical compounds as well as for exploring the mechanism of trans-translation in these pathogens.

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