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We recently identified the Pgf glycosylation machinery of <jats:italic>S. mutans<\/jats:italic>, responsible for the post\u2010translational modification of the surface\u2010associated adhesins Cnm and WapA. Since the four\u2010gene <jats:italic>pgf<\/jats:italic> operon (<jats:italic>pgfS<\/jats:italic>\u2010<jats:italic>pgfM1<\/jats:italic>\u2010<jats:italic>pgfE<\/jats:italic>\u2010<jats:italic>pgfM2<\/jats:italic>) is part of the <jats:italic>S. mutans<\/jats:italic> 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\u2010glucose polysaccharide synthesis pathway. Phenotypic characterization of <jats:italic>pgf<\/jats:italic> mutants (\u0394<jats:italic>pgfS<\/jats:italic>, \u0394<jats:italic>pgfE<\/jats:italic>, \u0394<jats:italic>pgfM1<\/jats:italic>, \u0394<jats:italic>pgfM2<\/jats:italic>, and \u0394<jats:italic>pgf<\/jats:italic>) revealed that the Pgf system is important for biofilm formation, surface charge, membrane stability, and survival in human saliva. Moreover, deletion of the entire <jats:italic>pgf<\/jats:italic> operon (\u0394<jats:italic>pgf<\/jats:italic> 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\u2010acetyl hexosamines but it becomes heavily phosphorylated with the inactivation of the PgfS glycosyltransferase, suggesting a crosstalk between these two post\u2010translational modification mechanisms. Our results revealed that the Pgf machinery contributes to multiple aspects of <jats:italic>S. mutans<\/jats:italic> pathobiology that may go beyond Cnm and WapA glycosylation.<\/jats:p>","DOI":"10.1111\/mmi.15190","type":"journal-article","created":{"date-parts":[[2023,11,17]],"date-time":"2023-11-17T08:17:10Z","timestamp":1700209030000},"page":"133-151","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Post\u2010translational modification by the Pgf glycosylation machinery modulates <i>Streptococcus mutans<\/i>\u00a0<scp>OMZ175<\/scp> physiology and virulence"],"prefix":"10.1111","volume":"122","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1194-0754","authenticated-orcid":false,"given":"Nicholas","family":"de Mojana\u00a0di\u00a0Cologna","sequence":"first","affiliation":[{"name":"Department of Oral Biology University of Florida, College of Dentistry  Gainesville Florida 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