Pleiotropic pigmentation mutants of Porphyromonas gingivalis

Porphyromonas gingivalis is a Gram-negative, black pigmented oral anaerobe associated with adult periodontitis. The adherence of the bacterium to junctional epithelial cells is the first step in infection and colonization. The molecular mechanisms and genetics of colonization are, as yet, not well understood, although it has been demonstrated that P. gingivalis fimbriae are involved in adhesion. In addition, cell surface cysteine proteinases may play a role either directly as adhesins or indirectly through their involvement in the biogenesis of fimbriae. A link has been established between cysteine proteinase-hemagglutinating activity and colongy pigmentation on blood agar. in this study a P. gingivalis ATCC 33277 transposon library was screened for white mutants. Pleiotropic mutants were identified with altered pigmentation, proteinase, hemagglutinin and haemolytic activities. Although the mutants fell into two classes based on the above phenotypes, by electron microscopy both classes showed increased fimbriation and decreased vesicle formation. Sequencing of genomic DNA flanking the transposon insertions revealed that one class of mutants carried disruptions in the gene encoding Lys-gingipain (kgp) and the other in a gene homologous to a glycosyl transferase. Potential roles for these genes in pigmentation, fimbriation, vesicle formation and attachment to epithelial cells are discussed. (C) 2000 Academic Press.