TraDIS-Xpress: a high-resolution whole-genome assay identifies novel mechanisms of triclosan action and resistance

Yasir, Muhammad, Turner, A. Keith, Bastkowski, Sarah, Baker, David, Page, Andrew J., Telatin, Andrea, Phan, Minh-Duy, Monahan, Leigh, Savva, George M., Darling, Aaron, Webber, Mark A. and Charles, Ian G. (2020) TraDIS-Xpress: a high-resolution whole-genome assay identifies novel mechanisms of triclosan action and resistance. Genome Research, 30 (2). pp. 239-249. ISSN 1088-9051

[img]
Preview
PDF (Published_Version) - Published Version
Available under License Creative Commons Attribution.

Download (8MB) | Preview

Abstract

Understanding the genetic basis for a phenotype is a central goal in biological research. Much has been learnt about bacterial genomes by creating large mutant libraries and looking for conditionally important genes. However, current genome-wide methods are largely unable to assay essential genes which are not amenable to disruption. To overcome this limitation, we developed a new version of “TraDIS” (transposon directed insertion-site sequencing) that we term “TraDIS-Xpress” that combines an inducible promoter into the transposon cassette. This allows controlled overexpression and repression of all genes owing to saturation of inserts adjacent to all open reading frames as well as conventional inactivation. We applied TraDIS-Xpress to identify responses to the biocide triclosan across a range of concentrations. Triclosan is endemic in modern life, but there is uncertainty about its mode of action with a concentration-dependent switch from bacteriostatic to bactericidal action unexplained. Our results show a concentration-dependent response to triclosan with different genes important in survival between static and cidal exposures. These genes include those previously reported to have a role in triclosan resistance as well as a new set of genes, including essential genes. Novel genes identified as being sensitive to triclosan exposure include those involved in barrier function, small molecule uptake, and integrity of transcription and translation. We anticipate the approach we show here, by allowing comparisons across multiple experimental conditions of TraDIS data, and including essential genes, will be a starting point for future work examining how different drug conditions impact bacterial survival mechanisms.

Item Type: Article
Additional Information: © 2020 Yasir et al.; Published by Cold Spring Harbor Laboratory Press.
Uncontrolled Keywords: bacteria,discovery,escherichia-coli,package
Faculty \ School: Faculty of Science > School of Biological Sciences
Faculty of Science > School of Computing Sciences
Faculty of Medicine and Health Sciences > Norwich Medical School
Faculty of Medicine and Health Sciences > School of Health Sciences
Related URLs:
Depositing User: LivePure Connector
Date Deposited: 20 Feb 2020 07:58
Last Modified: 28 Aug 2020 00:00
URI: https://ueaeprints.uea.ac.uk/id/eprint/74265
DOI: 10.1101/gr.254391.119

Actions (login required)

View Item View Item