Recombinant yeast and human cells as screening tools to search for antibacterial agents targeting the transcription termination factor Rho

Abstract : The alarming issue of antibiotic resistance expansion requires a continuous search for new and efficient antibacterial agents. Here we describe the design of new tools to screen for target-specific inhibitors of the bacterial Rho factor directly inside eukaryotic cells. Rho factor is a global regulator of gene expression which is essential to most bacteria, especially Gram-negative. Since Rho has no functional or structural homolog in eukaryotes, it constitutes a valuable and well known bacterial target as evidenced by its inhibition by the natural antibiotic, Bicyclomycin. Our screening tools are based on perturbation of mRNA processing and packaging reactions in the nucleus of eukaryotic cells by the RNA-dependent helicase/translocase activity of bacterial Rho factor leading to a growth defect phenotype. In this approach, any compound that impedes Rho activity should restore growth to yeast or human cells expressing Rho protein, providing valuable means to screen for target-specific antibacterial agents within the environment of a eukaryotic cell. The yeast tool expressing E. coli Rho factor was validated using Bicyclomycin as the control antibacterial agent. The validation of the screening tool was further extended with a stable human cell line expressing Rho factor conditionally. Finally, we show that Rho factors from different bacterial pathogens can also be designed as yeast-based screening tools which can reveal subtle variations in the functional features of the proteins.
Document type :
Journal articles
Complete list of metadatas

https://hal.archives-ouvertes.fr/hal-01966171
Contributor : Isabelle Frapart <>
Submitted on : Thursday, December 27, 2018 - 5:03:54 PM
Last modification on : Monday, May 13, 2019 - 12:34:08 PM

Identifiers

  • HAL Id : hal-01966171, version 1

Collections

Citation

Kévin Moreau, Justine Surand, Aurélia Le Dantec, Christine Mosrin-Huaman, Alain Legrand, et al.. Recombinant yeast and human cells as screening tools to search for antibacterial agents targeting the transcription termination factor Rho. Journal of Antibiotics, Nature Publishing Group: Open Access Hybrid Model Option B, 2018, 71 (4), pp.447-455. ⟨hal-01966171⟩

Share

Metrics

Record views

21