Acquired CRISPR spacers and rhamnose-glucose polysaccharide defects confer resistance to Streptococcus mutans phage ΦAPCM01
Streptococcus mutans is a major cause of dental caries worldwide. Targeted therapeutic strategies to eradicate S. mutans include oral phage rinses. In this study, we investigated how phage resistance develops in S. mutans. As a model phage, we used {phi}APCM01, which is known to infect a serotype e strain. We isolated and sequenced the genomes of 15 spontaneous resistant mutants and found that 10 had acquired novel CRISPR spacers targeting the phage, with a total of 18 new spacers identified. Additionally, eight strains contained mutations in rhamnose-glucose polysaccharide (RGP) biosynthetic genes, three of which also acquired spacers. Only the rgp mutants exhibited defects in phage absorption, supporting the role of these cell surface glycans as the phage receptor. Mutations in rgpF and the newly identified gene rgpX led to severe cell division defects and impaired biofilm formation, the latter of which shared by the rgpD mutant. Thus, rgp mutations confer phage resistance but impose severe fitness costs, limiting pathogenic potential. Surprisingly, we found that {phi}APCM01 was capable of binding to and injecting its genome into UA159, a model serotype c strain. However, UA159 was resistant to infection due to an unknown post-entry defense mechanism. Consequently, {phi}APCM01 has the potential to infect both major serotypes associated with dental caries. RepositoriesThe genome sequence of Streptococcus mutans DPC6143 was deposited at NCBI with the accession number NZ_CP172847.1.