Molecular and genetic analysis of multiple changes in the levels of production of virulence factors in a subcultured variant of Streptococcus mutans.

We previously isolated a variant strain, Xc100L, which shows decreased production of a surface protein antigen with a molecular mass of 190 kDa (PAc), after repeated subculturing of Streptococcus mutans strain Xc [Koga, T. et al. (1989) J.Gen. Microbiol. 135, 3199-3202]. In the present study, the levels of expression of the gtfB, gtfC, gtfD and ftf genes coding for polysaccharide-synthesizing enzymes in strain Xc100L were compared with those in strain Xc. Western blot analysis revealed multiple differences in the levels of production of these enzymes between these two strains. The amounts of the gtfB and gtfC gene products responsible for water-insoluble glucan synthesis in strain Xc100L were lower than those in strain Xc, whereas the amounts of the gtfD and ftf gene products responsible for water soluble glucan synthesis and fructan synthesis, respectively, in strain Xc100L were higher than those in strain Xc. Northern blot analysis revealed that the amounts of the four enzymes and PAc produced by strain Xc100L reflected the relative amounts of mRNAs from the genes. The chloramphenicol acetyltransferase gene was fused with each of these five genes, and the transcriptional activity of each gene in strain Xc100L was quantitatively compared with that in strain Xc. The chloramphenicol acetyltransferase assay also indicated that the phenotypic differences between strain Xc and strain Xc100L were due to differences in the transcriptional activities of the virulence genes. No differences in the nucleotide sequences of the promoter regions of the gtfB, gtfC, gtfD, ftf and pac genes were found between strain Xc and strain Xc100L. It is possible that a factor(s) affecting the levels of transcription of the multiple virulence genes exists in S. mutans.