The Pseudomonas aeruginosa global regulator MvaT specifically binds to the ptxS upstream region and enhances ptxS expression

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Abstract
Exotoxin A production inPseudomonas aeruginosais regulated positively or negatively by several genes. Two such regulatory genes,ptxRandptxS, which are divergently transcribed from each other, have been described previously. While computer analysis suggested that theptxR-ptxSintergenic region contains potential binding sites for several regulatory proteins, the mechanism that regulates the expression of eitherptxRorptxSinP. aeruginosais not known. The presence of aP. aeruginosaprotein complex that specifically binds to a segment within this region was determined. In this study the binding region was localized to a 150 bp fragment of the intergenic region and the proteins that constitute the binding complex were characterized asP. aeruginosaHU and MvaT. Recombinant MvaT was purified as a fusion protein (MAL-MvaT) and shown to specifically bind to theptxR-ptxSintergenic region. A PAO1 isogenic mutant defective inmvaT, PAOΔmvaT, was constructed and characterized. The lysate of PAOΔmvaTfailed to bind to the 150 bp probe. The effect ofmvaTonptxSandptxRexpression was examined using real-time PCR experiments. The expression ofptxSwas lower in PAOΔmvaTthan in PAO1, but no difference was detected inptxRexpression. These results suggest that MvaT positively regulatesptxSexpression by binding specifically to theptxSupstream region.