1887

Abstract

is a Gram-negative black-pigmented obligate anaerobe implicated in the aetiology of human periodontal disease. The virulence of is associated with the elaboration of the cysteine proteases Arg-gingipain (Rgp) and Lys-gingipain (Kgp), which are produced at high bacterial cell densities. To determine whether quorum sensing plays a role in the regulation of Rgp and Kgp, biosensors capable of detecting either -acylhomoserine lactone (AHLs) or the -dependent autoinducer (AI-2) quorum-sensing signalling molecules in spent culture supernatants were first employed. While no AHLs could be detected, the BB170 biosensor was activated by spent W50 culture supernatants. The gene was cloned and demonstrated to restore AI-2 production in the mutant DH5α. Mutation of abolished AI-2 production in . Western blotting using antibodies raised against the recombinant protein revealed that LuxS levels increased throughout growth even though AI-2 activity was only maximally detected at the mid-exponential phase of growth and disappeared by the onset of stationary phase. Similar results were obtained with DH5α transformed with , suggesting that AI-2 production is not limited by a lack of LuxS protein. Analysis of Rgp and Kgp protease activities revealed that the mutant produced around 45% less Rgp and 30% less Kgp activity than the parent strain. In addition, the mutant exhibited a fourfold reduction in haemagglutinin titre. However, these reductions in virulence determinant levels were insufficient to attenuate the mutant in a murine lesion model of infection.

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2002-03-01
2020-07-04
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