Curation Information

Publication
Transcriptome profiling defines a novel regulon modulated by the LysR-type transcriptional regulator MexT in Pseudomonas aeruginosa.;Tian ZX, Fargier E, Mac Aogáin M, Adams C, Wang YP, O'Gara F;Nucleic acids research 2009 Dec; 37(22):7546-59 [19846594]
TF
MexT [Q9I0Z0, view regulon]
Reported TF sp.
Pseudomonas aeruginosa PAO1
Reported site sp.
Pseudomonas aeruginosa PAO1
Created by
Dinara Sagitova
Curation notes
-

Experimental Process

A putative MexT regulon was identified by carrying out a whole-genome transcriptome profiling on PAO1 wild-type and an isogenic mexEF mutant overexpressing MexT. The validity of the microarray data was confirmed by a semi-quantitative reverse transcription PCR analysis of genes whose expression was significantly altered (PA4881, pqsA and lldP). A well-conserved DNA motif ATCA-N5-GTCGAT-N4-ACYAT was identified in the upstream regions of 10 genes by using MEME Suite online software. The transcriptional activation of the putative target genes by MexT was confirmed by constructing promoter-lacZ fusions of selected genes. To further investigate if these genes are direct targets of MexT, the ability of MexT to activate gene expression in a heterogeneous E. coli background was assessed. The results indicate that the conserved DNA motif is required for the induced expression of these genes by MexT. EMSAs and the site-directed mutagenesis showed that MexT protein bound to the conserved DNA motif and that a mutation in two of the most conserved nucleotides in the motif abolished the binding of the MexT protein. MexT binding to the conserved motif was abolished when a mutation in the helix–turn–helix motifwas introduced.

Transcription Factor Binding Sites


ATCATTTCTGTCGATAGTTAATAG
ATCAACCTTATCGATAAGTACCAT
ATCACCGCCGTCGATCTCGACTAA
ATCAGCCCTGTCGATAGCGACTAT
ATCATTTCTGTCGATAGTTAATAG
ATCAACCTTATCGATAAGTACCAT
ATCACCGCCGTCGATCTCGACTAA
ATCAGCCCTGTCGATAGCGACTAT

Gene Regulation

Regulated genes for each binding site are displayed below. Gene regulation diagrams show binding sites, positively-regulated genes, negatively-regulated genes, both positively and negatively regulated genes, genes with unspecified type of regulation. For each indvidual site, experimental techniques used to determine the site are also given.

Site sequence Regulated genes Gene diagram Experimental techniques TF function TF type
ATCATTTCTGTCGATAGTTAATAG PA1744,
... ... PA1744 PA1743
Experimental technique details Beta-gal reporter assay - Experimental technique details DNA-array expression analysis (ECO:0005525) - Experimental technique details Motif-discovery (ECO:0005558) - activator not specified
ATCAACCTTATCGATAAGTACCAT PA2759,
... ... PA2759 PA2760 PA2761
Experimental technique details Beta-gal reporter assay - Experimental technique details DNA-array expression analysis (ECO:0005525) - Experimental technique details Motif-discovery (ECO:0005558) - activator not specified
ATCACCGCCGTCGATCTCGACTAA PA3229
... ... PA3229
Experimental technique details Beta-gal reporter assay - Experimental technique details DNA-array expression analysis (ECO:0005525) - Experimental technique details Motif-discovery (ECO:0005558) - activator not specified
ATCAGCCCTGTCGATAGCGACTAT PA4354,
... ... PA4354 PA4353 PA4352 PA4355 xenB
Experimental technique details Beta-gal reporter assay - Experimental technique details DNA-array expression analysis (ECO:0005525) - Experimental technique details Motif-discovery (ECO:0005558) - activator not specified