1887

Abstract

The HpmA haemolysin toxin of is encoded by the locus and its production is upregulated co-ordinately with the synthesis and assembly of flagella during differentiation into hyperflagellated swarm cells. Primer extension identified a σ promoter upstream of that was upregulated during swarming. Northern blotting indicated that this promoter region was also required for concomitant transcription of the immediately distal gene, and that the unstable transcript generated a stable mRNA and an unstable mRNA. Transcriptional fusions to the DNA regions 5′ of the and genes confirmed that σ promoter activity increased in swarm cells, and that there was no independent promoter. Increased transcription of the operon in swarm cells was dependent upon a 125 bp sequence 5′ of the σ promoter −35 hexamer. This sequence spans multiple putative binding sites for the leucine-responsive regulatory protein (Lrp), and band-shift assays with purified Lrp confirmed the presence of at least two such sites. The influence on expression of the key swarming positive regulators FlhDC (encoded by the flagellar master operon), Lrp, and the membrane-located upregulator of the master operon, UmoB, was examined. Overexpression of each of these regulators moderately increased transcription in wild-type , and the operon was not expressed in any of the , or mutants. Expression in the mutants was not recovered by cross-complementation, i.e. by overexpression of FlhDC, Lrp or UmoB. Expression of the protease virulence gene, which like is also upregulated in swarm cells, did not require Lrp, but like it was upregulated by UmoB. The results indicate intersecting pathways of control linking virulence gene expression and swarm cell differentiation.

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2002-07-01
2019-09-16
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