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bioRxiv · 10.1101/2020.12.16.423090

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

Thepaut, M.·Campos Da Silva, R.·Renard, E.·Barloy-Hubler, F.·Ennifar, E.·Boujard, D.·Gillet, R.
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Abstract

Bacteria cope with ribosome stalling thanks to trans-translation, a major quality control system of protein synthesis that is mediated by tmRNA, an hybrid RNA with properties of both a tRNA and an 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, none of these permits the safe and easy evaluation of trans-translation in 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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Thepaut, M., Campos Da Silva, R., Renard, E., Barloy-Hubler, F., Ennifar, E., Boujard, D., Gillet, R.. 2020-12-17. Safe and easy evaluation of tmRNA-SmpB-mediated trans-translation in ESKAPE pathogenic bacteria. https://doi.org/10.1101/2020.12.16.423090

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