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Nucleic Acids Research Advance Access published online on October 21, 2009
Nucleic Acids Research, doi:10.1093/nar/gkp828
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© The Author(s) 2009. Published by Oxford University Press.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Molecular Biology

Transcriptome profiling defines a novel regulon modulated by the LysR-type transcriptional regulator MexT in Pseudomonas aeruginosa

Zhe-Xian Tian1,2, Emilie Fargier1, Micheál Mac Aogáin1, Claire Adams1, Yi-Ping Wang2 and Fergal O’Gara1,*

1BIOMERIT Research Centre, Department of Microbiology, University College Cork, Cork, Ireland and 2National Laboratory of Plant Engineering and Protein Genetic Engineering, College of Life Science, Peking University, Beijing, P.R. China

*To whom correspondence should be addressed. Tel: +353 21 4902861; Fax: +353 21 4903101; Email: f.ogara{at}ucc.ie

Received July 22, 2009. Revised September 18, 2009. Accepted September 18, 2009.

The LysR-family regulator MexT modulates the expression of the MexEF-OprN efflux system in the human pathogen Pseudomonas aeruginosa. Recently, we demonstrated that MexT regulates certain virulence phenotypes, including the type-three secretion system and early attachment independent of its role in regulating MexEF-OprN. In this study, transcriptome profiling was utilized to investigate the global nature of MexT regulation in P. aeruginosa PAO1 and an isogenic mexEF mutant. Twelve genes of unknown function were highly induced by overexpressing MexT independent of MexEF-OprN. A well-conserved DNA motif was identified in the upstream regulatory region of nine of these genes and upstream of mexE. Reporter fusion analysis demonstrated that the expression of the genes was significantly induced by MexT in P. aeruginosa and a heterogenous Escherichia coli strain and that the conserved sequence was required for this induction. The conserved DNA motif was further characterized as the MexT binding site by site-directed mutagenesis and electrophoretic mobility shift assays. Genes containing this conserved regulatory sequence were identified across other Pseudomonas species, and their expression was activated by MexT. Thus, a novel regulon directly modulated by MexT, that includes but is independent of mexEF-oprN, has been identified.


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