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The causative agent of ovine footrot, the gram-negative anaerobeDichelobacter nodosus , produces polar type IV fimbriae, which are the major protective antigens. TheD. nodosus genesfimN ,fimO , andfimP are homologs of thePseudomonas aeruginosa fimbrial assembly genes,pilB ,pilC , andpilD , respectively. Both thepilD andfimP genes encode prepilin peptidases that are responsible for cleavage of the leader sequence from the immature fimbrial subunit. To investigate the functional similarity of the fimbrial biogenesis systems from these organisms, theD. nodosus genes were introduced intoP. aeruginosa strains carrying mutations in the homologous genes. Analysis of the resultant derivatives showed that thefimP gene complemented apilD mutant ofP. aeruginosa for both fimbrial assembly and protein secretion. However, thefimN andfimO genes did not complementpilB orpilC mutants, respectively. These results suggest that although the PilD prepilin peptidase can be functionally replaced by the heterologous FimP protein, the function of the PilB and PilC proteins may require binding or catalytic domains specific for theP. aeruginosa fimbrial assembly system. The transcriptional organization and regulation of thefimNOP gene region were also examined. The results of reverse transcriptase PCR and primer extension analysis suggested that these genes form an operon transcribed from two ς70 -type promoters located upstream of ORFM, an open reading frame proximal tofimN . Transcription of theD. nodosus fimbrial subunit was found to increase in cells grown on solid media, and it was postulated that this regulatory effect may be of significance in the infected footrot lesion.

infection and immunity

Complementation Analysis of the Dichelobacter nodosus fimN , fimO , and fimP Genes in Pseudomonas aeruginosa and Transcriptional Analysis of the fimNOP Gene Region