Functional characterization of FlgM in the regulation of flagellar synthesis and motility in Yersinia pseudotuberculosis
Autor: | Steve Atkinson, Lisha Ding, Yangbo Hu, Yao Wang, Paul Williams, Shiyun Chen |
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Rok vydání: | 2009 |
Předmět: |
Transcription
Genetic Molecular Sequence Data Mutant Sigma Factor Flagellum Microbiology Bacterial Proteins Sigma factor Transcription (biology) Gene expression Yersinia pseudotuberculosis Amino Acid Sequence Amino Acids Promoter Regions Genetic Conserved Sequence Regulation of gene expression Genetics biology Escherichia coli Proteins Genetic Complementation Test Gene Expression Regulation Bacterial biology.organism_classification Flagella Mutagenesis Site-Directed biology.protein Flagellin Protein Binding |
Zdroj: | Microbiology. 155:1890-1900 |
ISSN: | 1465-2080 1350-0872 |
Popis: | We describe here the functional characterization of the flgM gene in Yersinia pseudotuberculosis. Direct interaction of FlgM with the alternative sigma factor σ 28 (FliA) was first confirmed. A conserved region in the C-terminus of FlgM was found which included the σ 28 binding domain. By site-directed mutagenesis, bacterial two-hybrid analysis and Western blotting, the primary FlgM binding sites with σ 28 were shown to be Ile85, Ala86 and Leu89. A role for FlgM in swimming motility was demonstrated by inactivation of flgM and subsequent complementation in trans. Transcriptional fusion analyses showed differential gene expression of flhDC, fliA, flgM and fliC in the fliA and flgM mutants compared with the wild-type. flhDC expression was not influenced by σ 28 or FlgM while fliA expression was abolished in the fliA mutant and considerably reduced in the flgM mutant when compared to the wild-type, indicating that both FliA and FlgM can activate fliA transcription. Conversely, flgM transcription was higher in the fliA mutant when compared to the wild-type, suggesting that flgM transcription was repressed by σ 28. Interestingly, fliC expression was markedly increased in the flgM mutant, suggesting a negative regulatory role for FlgM in fliC expression. The transcription of other σ-dependent genes (cheW, flgD, flaA, csrA and fliZ) was also examined in fliA and flgM mutant backgrounds and this revealed that other σ-factors apart from σ 28 may be involved in flagellar biogenesis in Y. pseudotuberculosis. Taking together the motility phenotypes and effects of flgM mutation on the regulation of these key motility genes, we propose that the mechanisms regulating flagellar biogenesis in Y. pseudotuberculosis may differ from those described for other bacteria. |
Databáze: | OpenAIRE |
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