Analysis of the virulence of Streptococcus mutans serotype c gtfA mutants in the rat model system.

The Streptococcus mutans serotype c gtfA gene encodes a 55-kilodalton protein which catalyzes the synthesis of a small glucan (1.5 kilodaltons) from sucrose (J.P. Robeson, R.G. Barletta, and R. Curtiss III, J. Bacteriol. 153:211-221, 1983). To investigate the role of the GtfA enzyme in virulence, we constructed S. mutans gtfA mutants from three cariogenic serotype c strains. A plasmid that carried an erythromycin resistance determinant and an internal fragment of the gtfA gene but that was unable to replicate in streptococci was used to transform S. mutans. The erythromycin-resistant transformants carried a partial duplication of the internal gtfA fragment, because of the integration of plasmid sequences within the S. mutans gtfA gene, which also resulted in the inactivation of the gtfA gene. This was verified by Southern DNA hybridization analysis and Western blot studies of cellular protein extracts of the mutant strains with GtfA antiserum. Mutants were fully virulent in both germfree and conventional rats. These results do not rule out the involvement of the GtfA protein in virulence. Pucci and Macrina (M.J. Pucci and F.L. Macrina, Infect. Immun. 54:77-84, 1986) have suggested that the GtfA enzyme synthesizes a primer for water-insoluble glucans. Another S. mutans protein, presumably a glucosyltransferase, may have a similar function and, thus, may obscure the relevance of the GtfA enzyme in pathogenesis.