Novel genes that upregulate the Proteus mirabilis flhDC master operon controlling flagellar biogenesis and swarming.
By screening for restoration of multicellular migration in a non-swarming but motile Proteus mirabilis mutant lacking the FIgN facilitator of flagella assembly, we identified four distinct genes that, in trans and multicopy, increased flagella production and cell length. Each of the genes upregulated expression of the flhDC master operon that controls flagellar biogenesis, cell division and swarming, not only in the mutant but also in the wild type. The genes were named umoA, umoB, umoC and umoD. Disruption of each of the wild-type chromosomal umo genes caused corresponding reductions in swarming and cell elongation, which correlated with decreased expression of the flhDC operon. The umoA, umoB, umoC and umoD genes are not closely linked, and only umoB is part of an operon. The sequences of the calculated gene products, UmoA (20.6 kDa), UmoB (78.0 kDa), UmoC (15.2 kDa) and UmoD (19.2 kDa), contain putative N-terminal secretion signals and predict a location in the cell membranes or periplasm. UmoB and UmoD have sequence similarity to the Escherichia coli uncharacterized open reading frames YrfF and YcfJ respectively; UmoA and UmoC have no known homologues. The umoB and umoC gene transcripts were present at very low levels, but umoA and umoD expression was similar to that of flhDC and increased in parallel with flhDC expression during differentiation into elongated hyperflagellated swarm cells. Like flhDC, umoA and umoD expression was subject to negative feedback in aflagellar assembly mutant lacking the FlhA inner membrane component of the export machinery. Assays of umo gene expression and cross-complementation indicated that the umo genes do not act in sequence within a pathway to upregulate flhDC, but revealed that umoA and umoD are reciprocally upregulated by FlhDC. Our findings strengthen the picture of the flhDC master operon as a major assimilatory checkpoint in Proteus mirabilis and other Gram-negative bacteria and expand the view of a complex regulatory network coupled to flagellar biogenesis.
University of Cambridge Department of Pathology, UK.
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Dufour
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Furness
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Hughes
D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D002876:Chromosomes, Bacterial; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins / Q000235:genetics*; D029968:Escherichia coli Proteins; D005407:Flagella / Q000502:physiology*; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D008565:Membrane Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D009876:Operon; D011513:Proteus mirabilis / Q000235:genetics* / Q000502:physiology; D015534:Trans-Activators / Q000235:genetics*; D015854:Up-Regulation
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C080688:FlhA protein, Bacteria; D008565:Membrane Proteins; D015534:Trans-Activators; C090552:flhC protein, E coli; C090553:flhD protein, E coli