1887

Abstract

is a Gram-negative pathogen that causes enteric redmouth disease in salmonids. A gene from encoding an extracellular protease termed ( rotease ) was cloned from a 3AI library constructed in pUC19 and analysed in gelatin-supplemented medium. The nucleotide sequence of the gene indicated an ORF encoding a protein of 477 aa. On the basis of the high degree of homology in the amino acid sequence as well as its conservative motifs, this protein was included within the serralysin metalloendopeptidase subfamily (EC 3.4.24.12). The N-terminal sequence showed a 14 aa propeptide followed by a 10 aa sequence identical to the one deduced previously from the 47 kDa purified protease. Additional results demonstrated that the gene encodes the 47 kDa protein. In contrast to other species, the protease is secreted by a type I Gram-negative bacterial ABC exporter protein secretion system composed of three genes termed , and , and a protease inhibitor . The development of genetic methods for this species has allowed the exploration of the organization and the putative role of the Yrp1 genetic locus. Thus, site-directed insertion mutations into the and the genes were constructed by the integration of the mobilizable suicide vector pIVET8 containing internal portions of both coding sequences. Complementation studies of those mutants with different loci indicated that they are organized as a single operon. The mutant strains lacked protease activity as well as the Yrp1 protein and, although physiologically similar to the parental strain when growing on nutrient broth medium, they were attenuated in virulence when bacteria were injected intraperitoneally into rainbow trout (). This is the first report of defined mutations in to show the implication of a factor such as an extracellular protease in pathogenesis.

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2002-07-01
2020-09-23
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