1887

Abstract

We describe here the functional characterization of the gene in . Direct interaction of FlgM with the alternative sigma factor (FliA) was first confirmed. A conserved region in the C-terminus of FlgM was found which included the binding domain. By site-directed mutagenesis, bacterial two-hybrid analysis and Western blotting, the primary FlgM binding sites with were shown to be Ile85, Ala86 and Leu89. A role for FlgM in swimming motility was demonstrated by inactivation of and subsequent complementation . Transcriptional fusion analyses showed differential gene expression of , , and in the and mutants compared with the wild-type. expression was not influenced by or FlgM while expression was abolished in the mutant and considerably reduced in the mutant when compared to the wild-type, indicating that both FliA and FlgM can activate transcription. Conversely, transcription was higher in the mutant when compared to the wild-type, suggesting that transcription was repressed by . Interestingly, expression was markedly increased in the mutant, suggesting a negative regulatory role for FlgM in expression. The transcription of other -dependent genes (, , , and ) was also examined in and mutant backgrounds and this revealed that other -factors apart from may be involved in flagellar biogenesis in . Taking together the motility phenotypes and effects of mutation on the regulation of these key motility genes, we propose that the mechanisms regulating flagellar biogenesis in may differ from those described for other bacteria.

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2009-06-01
2020-04-05
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