A motile but non-swarming mutant of Proteus mirabilis lacks FlgN, a facilitator of flagella filament assembly.
A TnphoA mutant of Proteus mirabilis was isolated, which had lost the ability to swarm, yet was still motile. The transposon had inserted into flgN, a flagella gene encoding a 147-amino-acid protein of undefined function. Proteus flgN is arranged in an operon with the class III anti-sigma28 gene, flgM, flanked by the class II genes, flgA, flgBCD and flhBA, and a novel putative virulence-related gene. The flgN mutation caused a substantial reduction in cell surface-associated flagellin, particularly during differentiation to the normally hyperflagellated swarm cell. This was not due to an effect on flagella gene expression or a typical defect in the flagella export apparatus as there was no class III gene downregulation by FlgM feedback, or intracellular flagellin accumulation. Loss of FlgN nevertheless caused a severe reduction in the incorporation of pulse-labelled flagellin into the membrane/flagellum fraction of differentiating cells. Substantial amounts of both non-oligomeric flagellin and flagellin degradation products appeared in the extracellular medium, although the few mature filaments made by the mutant were no more sensitive to proteolysis than those of the wild type. FlgN appeared soluble and active in the cytosol. The data suggest that the function of FlgN is to facilitate the initiation of flagella filament assembly, a role that may be especially critical in attaining the much higher concentration of surface flagellin required for swarming. Proteus FlgN has leucine zipper-like motifs arranged on potential amphipathic helices, a feature conserved in cytosolic chaperones for the exported substrates of flagella-related type III virulence systems. While gel filtration of FlgN from the soluble cell fraction did not establish an interaction with flagellin, it indicated that FlgN may associate with an unknown component and/or form an oligomer.
Gygi
D
D
University of Cambridge Department of Pathology, UK.
Fraser
G
G
Dufour
A
A
Hughes
C
C
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology*; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004251:DNA Transposable Elements / Q000235:genetics; D005407:Flagella / Q000235:genetics* / Q000502:physiology*; D005408:Flagellin / Q000235:genetics / Q000378:metabolism; D005798:Genes, Bacterial; D018832:Molecular Chaperones / Q000737:chemistry; D008969:Molecular Sequence Data; D009068:Movement; D009154:Mutation; D011513:Proteus mirabilis / Q000235:genetics* / Q000472:pathogenicity / Q000502:physiology*; D017386:Sequence Homology, Amino Acid; D012995:Solubility; D014774:Virulence
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; C087498:FlgN protein, Bacteria; D018832:Molecular Chaperones; D005408:Flagellin