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Identification of a novel gene, fimV, involved in twitching motility in Pseudomonas aeruginosa

Identification of a novel gene, fimV, involved in twitching motility in Pseudomonas aeruginosa,Annalese B. T. Semmler,Cynthia B. Whitchurch,Andrew J.

Identification of a novel gene, fimV, involved in twitching motility in Pseudomonas aeruginosa   (Citations: 17)
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Transposon mutagenesis was used to identify a new locus required for twitching motility in Pseudomonas aeruginosa. Four Tn5-B21 mutants which lacked twitching motility and a fifth which exhibited impaired motility were found to map to the same KpnI restriction fragment at approximately 40 min on the P. aeruginosa genome. Cloning and sequencing studies showed that all five transposon insertions occurred within the same 2<8 kb ORF, which was termed fimV. The product of this gene has a putative peptidoglycan-binding domain, predicted transmembrane domains, a highly acidic C terminus and anomalous electrophoretic migration, indicating unusual primary or secondary structure. The P. aeruginosa genome also possesses a paralogue of fimV. Homologues of fimV were also found in the sequenced genomes of the other type-IV-fimbriated bacteria Neisseria gonorrhoeae, Neisseria meningitidis, Legionella pneumophila and Vibrio cholerae, but not in those of other bacteria which lack type IV fimbriae. A fimV homologue was also found in the genome sequence of Shewanella putrefaciens, along with many other homologues of type IV fimbrial genes, indicating that this bacterium is also likely to produce type IV fimbriae. Wild-type twitching motility was restored to fimV mutants by complementation in a dosage-dependent manner. Overexpression of fimV resulted in an unusual phenotype where the cells were massively elongated and migrated in large convoys at the periphery of the colony. It is suggested that FimV may be involved in remodelling of the peptidoglycan layer to enable assembly of the type IV fimbrial structure and machinery.
Published in 2000.
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    • ...By transposon mutagenesis, genes essential for twitching motility in Pseudomonas aeruginosa were identified including fimV (Semmler et al. 2000; Ahn et al. 2004; Oldfield et al. 2007)...
    • ...Originally identified (Semmler et al. 2000) as conserved between L. pneumophila, Neisseria gonorrhoeae, Neisseria meningitidis, P. aeruginosa, and Shewanella putrefaciens, homologs have since been discovered in a wide range of other bacterial species which possess type IV pili...
    • ...While the function of this protein is not known, it has been observed that overexpression of FimV in P. aeruginosa causes a dramatic elongation of the cells and that the peptidoglycan-binding domain is conserved between species (Semmler et al. 2000)...
    • ...Therefore, it has been proposed to play a role in remodeling the peptidoglycan layer for insertion of the type IV pilus through the outer membrane and in maintaining cell morphology (Semmler et al. 2000)...
    • ...The inducible tac promoter (containing the lacI repressor) that is present in pMMBN is known to be ‘‘leaky’’ and to provide a basal level of transcription in the absence of induction with IPTG (e.g. Semmler et al. 2000)...
    • ...However, DfimV cells are quite abnormally long, similar to what was observed when fimV was overexpressed in P. aeruginosa (Semmler et al. 2000)...
    • ...Very little is known about the actual functioning of the fimV gene, beyond the hypothesis from P. aeruginosa that it might be involved in the remodeling of the peptidoglycan layer (Semmler et al. 2000)...

    David A. CoilJozefet al. The role of fimV and the importance of its tandem repeat copy number i...

    • ...LPG1299 is homologous to the fimV gene described in Pseudomonas aeruginosa [41]...

    David A Coilet al. Intragenic tandem repeat variation between Legionella pneumophila stra...

    • ...The level of intracellular pilin expression and the amount of surface pili of tonB mutants were analyzed with semiquantitative Western blotting and quantitative enzyme-linked immunosorbent assays (ELISAs) (1, 13), respectively, using rabbit anti-PAK pilus antibodies...

    Bixing Huanget al. tonB3 Is Required for Normal Twitching Motility and Extracellular Asse...

    • ...Western blotting to detect pili in the surface and whole-cell fractions and quantification of the surface pili in P. aeruginosa cultures by an enzyme-linked immunosorbent assay (ELISA) were carried out as described previously (3, 41)...
    • ...Light microscopy of twitching zones was performed as described previously (41)...
    • ...In addition, expression of cloned fimX did not impair twitching motility in wild-type cells, confirming that the activity of FimX was not dose sensitive (data not shown), in contrast to the activities of some other proteins which affect twitching motility, such as FimV (41)...
    • ... (Fig. 2G to L). Normal twitching motility in wild-type cells involves outward movement of broad rafts of cells, followed by a breaking up of the rafts into a thinner lattice-like network, within which cells traverse up and down with frequent reversals of movement (Fig. 2G), whereas pilA mutants (which lack type IV pili and twitching motility) have smooth and relatively static colony edges (Fig. 2H) and exhibit no network formation (41)...

    Bixing Huanget al. FimX, a Multidomain Protein Connecting Environmental Signals to Twitch...

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