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

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 regula...

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Bibliographic Details
Published in:	Journal of antibiotics Vol. 71; no. 4; pp. 447 - 455
Main Authors:	Moreau, Kévin, Surand, Justine, Le Dantec, Aurélia, Mosrin-Huaman, Christine, Legrand, Alain, Rahmouni, A. Rachid
Format:	Journal Article
Language:	English
Published:	London Nature Publishing Group UK 01-03-2018 Nature Publishing Group Nature Publishing Group: Open Access Hybrid Model Option B
Subjects:	38/44 631/337/572 692/699/255/1318 Anti-Bacterial Agents - pharmacology Antibacterial agents Antibiotic resistance Antibiotics Bacteria Bacterial Infections - microbiology Bacteriology Biomedical and Life Sciences Bioorganic Chemistry Cell Line Cell Survival - drug effects Drug Evaluation, Preclinical E coli Escherichia coli - drug effects Escherichia coli - genetics Gene expression Gram-Negative Bacteria - drug effects HEK293 Cells Humans Life Sciences Medicinal Chemistry Microbiology Organic Chemistry Rho Factor - drug effects Saccharomyces cerevisiae - drug effects Transcription, Genetic Yeasts Yeasts - drug effects
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Summary:	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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0021-8820 1881-1469
DOI:	10.1038/s41429-017-0007-x