Post-translational modification by the Pgf glycosylation machinery modulates Streptococcus mutans OMZ175 physiology and virulence

Streptococcus mutans is commonly associated with dental caries and the ability to form biofilms is essential for its pathogenicity. We recently identified the Pgf glycosylation machinery of S. mutans, responsible for the post-translational modification of the surface-associated adhesins Cnm and WapA. Since the four-gene pgf operon (pgfS-pgfM1-pgfE-pgfM2) is part of the S. mutans core genome, we hypothesized that the scope of the Pgf system goes beyond Cnm and WapA glycosylation. In silico analyses and tunicamycin sensitivity assays suggested a functional overlap between the Pgf machinery and the rhamnose-glucose polysaccharide synthesis pathway. Phenotypic characterization of pgf mutants (Delta pgfS, Delta pgfE, Delta pgfM1, Delta pgfM2, and Delta pgf) revealed that the Pgf system is important for biofilm formation, surface charge, membrane stability, and survival in human saliva. Moreover, deletion of the entire pgf operon (Delta pgf strain) resulted in significantly impaired colonization in a rat oral colonization model. Using Cnm as a model, we showed that Cnm is heavily modified with N-acetyl hexosamines but it becomes heavily phosphorylated with the inactivation of the PgfS glycosyltransferase, suggesting a crosstalk between these two post-translational modification mechanisms. Our results revealed that the Pgf machinery contributes to multiple aspects of S. mutans pathobiology that may go beyond Cnm and WapA glycosylation.

Automated summary

Streptococcus mutans is commonly associated with dental caries and the ability to form biofilms is essential for its pathogenicity. The study identified the Pgf glycosylation machinery of S. mutans and found that it is not only related to the glycosylation of Cnm and WapA, but also to other biosynthetic pathways. Phenotypic characterization of pgf mutants revealed that the Pgf system is important for biofilm formation, surface charge, membrane stability, and survival in human saliva. Deletion of the entire pgf operon resulted in significantly impaired colonization in a rat oral colonization model.