1887

Abstract

Flagellar biogenesis in the gastric pathogen involves a transcriptional hierarchy that utilizes all three sigma factors found in this bacterium (RpoD, RpoN and FliA). Transcription of the RpoN-dependent genes requires the sensor kinase FlgS and response regulator FlgR. It is thought that FlgS senses some cellular cue to regulate transcription of the RpoN-dependent flagellar genes, but this signal has yet to be identified. Previous studies showed that transcription of the RpoN-dependent genes is inhibited by mutations in , which encodes a membrane-bound component of the flagellar protein export apparatus. We found that depending on the strain used, insertion mutations in had different effects on expression of RpoN-dependent genes. Mutations in in strains B128 and ATCC 43504 (the type strain) were generated by inserting a chloramphenicol resistance cassette so as to effectively eliminate expression of the gene (Δ), or within the gene following codon 77 (designated ) or codon 454 (designated ), which could allow expression of truncated FlhA proteins. All three mutations severely inhibited transcription of the RpoN-dependent genes and in B128. In contrast, levels of and transcripts in ATCC 43504 bearing either or , but not Δ, were ~60 % of wild-type levels. The FlhA variant was detected in membrane fractions prepared from ATCC 43504 but not B128, which may account for the phenotypic differences in the mutations of the two strains. Taken together, these findings suggest that only the N-terminal region of FlhA is needed for transcription of the RpoN regulon. Interestingly, expression of an ′-′ reporter gene in ATCC 43504 bearing the allele was about eightfold higher than that of a strain with the wild-type allele, suggesting that expression of is not only regulated at the level of transcription but also regulated post-transcriptionally.

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2013-01-01
2020-01-23
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