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

Yasir, M; Turner, AK; Bastkowski, S; Baker, D; Page, AJ; Telatin, A; Phan, M-D; Monahan, L; Savva, GM; Darling, A; Webber, MA; Charles, IG

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

Yasir, M Turner, AK Bastkowski, S Baker, D Page, AJ Telatin, A Phan, M-D Monahan, L

Savva, GM Darling, A

Webber, MA Charles, IG

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Publisher:: Cold Spring Harbor Laboratory
Publication Type:: Journal Article
Citation:: Genome research, 2020, 30, (2), pp. 239-249
Issue Date:: 2020-02-12

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Field	Value	Language
dc.contributor.author	Yasir, M
dc.contributor.author	Turner, AK
dc.contributor.author	Bastkowski, S
dc.contributor.author	Baker, D
dc.contributor.author	Page, AJ
dc.contributor.author	Telatin, A
dc.contributor.author	Phan, M-D
dc.contributor.author	Monahan, L https://orcid.org/0000-0003-2933-9000
dc.contributor.author	Savva, GM
dc.contributor.author	Darling, A https://orcid.org/0000-0003-2397-7925
dc.contributor.author	Webber, MA
dc.contributor.author	Charles, IG
dc.date.accessioned	2020-08-14T06:29:51Z
dc.date.available	2020-01-17
dc.date.available	2020-08-14T06:29:51Z
dc.date.issued	2020-02-12
dc.identifier.citation	Genome research, 2020, 30, (2), pp. 239-249
dc.identifier.issn	1088-9051
dc.identifier.issn	1549-5469
dc.identifier.uri	http://hdl.handle.net/10453/142108
dc.description.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.
dc.format	Print-Electronic
dc.language	eng
dc.publisher	Cold Spring Harbor Laboratory
dc.relation.ispartof	Genome research
dc.relation.isbasedon	10.1101/gr.254391.119
dc.rights	info:eu-repo/semantics/openAccess
dc.subject	06 Biological Sciences, 11 Medical and Health Sciences
dc.subject.classification	Bioinformatics
dc.title	TraDIS-Xpress: a high-resolution whole-genome assay identifies novel mechanisms of triclosan action and resistance.
dc.type	Journal Article
utslib.citation.volume	30
utslib.location.activity	United States
utslib.for	06 Biological Sciences
utslib.for	11 Medical and Health Sciences
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Strength - ithree - Institute of Infection, Immunity and Innovation
pubs.organisational-group	/University of Technology Sydney
utslib.copyright.status	open_access	*
dc.date.updated	2020-08-14T06:28:58Z
pubs.issue	2
pubs.publication-status	Published
pubs.volume	30
utslib.citation.issue	2

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.

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http://hdl.handle.net/10453/142108