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FliT Acts as an Anti-FlhD₂C₂ Factor in the Transcriptional Control of the Flagellar Regulon in Salmonella enterica Serovar Typhimurium

Shouji Yamamoto and Kazuhiro Kutsukake^*

Graduate School of Natural Science and Technology and Department of Biology, Faculty of Science, Okayama University, Tsushima-Naka 3-1-1, Okayama 700-8530, Japan

Received 5 June 2006/ Accepted 7 July 2006

Flagellar operons are divided into three classes with respect to their transcriptional hierarchy in Salmonella enterica serovar Typhimurium. The class 1 gene products FlhD and FlhC act together in an FlhD₂C₂ heterotetramer, which binds upstream of the class 2 promoters to facilitate binding of RNA polymerase. Class 2 expression is known to be enhanced by a disruption mutation in a flagellar gene, fliT. In this study, we purified FliT protein in a His-tagged form and showed that the protein prevented binding of FlhD₂C₂ to the class 2 promoter and inhibited FlhD₂C₂-dependent transcription. Pull-down and far-Western blotting analyses revealed that the FliT protein was capable of binding to FlhD₂C₂ and FlhC and not to FlhD alone. We conclude that FliT acts as an anti-FlhD₂C₂ factor, which binds to FlhD₂C₂ through interaction with the FlhC subunit and inhibits its binding to the class 2 promoter.

Present address: Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Higashi-Hiroshima, Hiroshima 739-8530, Japan.

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