1887

Abstract

The role of the P.69/pertactin protein in mammalian cell adhesion and invasion was investigated. strains expressing surface-associated P.69/pertactin from a chromosomally located gene were significantly more invasive than isogenic parental strains. This effect was most pronounced in the poorly invasive, semi-rough strain LB5010. K-12 strain HB101 harbouring the plasmid p41869D, which encodes the full-length gene under the control of the promoter on the broad-host-range plasmid pMMB66EH, was significantly more adhesive to HEp-2 and Chinese Hamster Ovary (CHO) cells growing in culture than HB101(pMMB66EH). However, the ability of to invade mammalian cells was not affected by P.69/pertactin expression. P.69/pertactin-mediated adhesiveness of HB101 to HEp-2 and CHO cells was not influenced by the viability of the bacterial cells. However, adherence was markedly reduced when assays were performed for less than 3 h, at 4°C or in the presence of cycloheximide, suggesting the active participation of the eukaryotic cell in bacterial adhesion. Site-directed mutagenesis was used to mutate Asp to Glu in an Arg-Gly-Asp (RGD→RGE) sequence present in mature P.69/pertactin and the mutated gene was cloned in the same broad-host-range vector (plasmid p41869E). This mutation had no detectable influence on the ability of P.69/pertactin to mediate adhesion of HB101 to HEp-2 or CHO cells. Plasmids p41869D and p41869E were introduced into the -negative strain BP347. Expression of P.69RGD or P.69RGE did not enhance the adhesiveness of BP347 for epithelial (HEp-2 and CHO) cells.

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1996-11-01
2021-05-12
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