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Volume 171, Issue 6

Acquired CRISPR spacers and rhamnose-glucose polysaccharide defects confer resistance to phage ɸAPCM01 Open Access

Abstract

is commonly associated with the development of dental caries worldwide. Due to their specificity for , phage represents a promising avenue for future targeted therapeutic strategies. In this study, we investigated how phage resistance develops in . As a model phage, we used ɸAPCM01, 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 clustered regularly interspaced short palindromic repeats (CRIPSR) spacers targeting the phage, with a total of 18 new spacers identified. Additionally, eight strains contained mutations in rhamnose-glucose polysaccharide biosynthetic genes, three of which also acquired spacers. Only the mutants exhibited defects in phage adsorption, supporting the role of these cell surface glycans as the phage receptor. Mutations in and the newly identified gene led to severe cell division defects and impaired biofilm formation, the latter of which was also shared by an mutant. Thus, mutations confer phage resistance but impose severe fitness costs, limiting pathogenic potential. Surprisingly, we found that ɸAPCM01 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 defence mechanism. Consequently, ɸAPCM01 has the potential to infect both major serotypes associated with dental caries.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): bacteriophage , clustered regularly interspaced short palindromic repeats (CRISPR)-Cas , dental caries , resistance , rhamnose-glucose polysaccharide and Streptococcus mutans
Funding
This study was supported by the:
  • National Institute of General Medical Sciences (Award P20GM103432)
    • Principal Award Recipient: LucasA. Wall
  • National Institute of General Medical Sciences (Award GM140886)
    • Principal Award Recipient: DanielWall
© 2025 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
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2025-06-19
2026-04-15

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/content/journal/micro/10.1099/mic.0.001575
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Acquired CRISPR spacers and rhamnose-glucose polysaccharide defects confer resistance to Streptococcus mutans phage ɸAPCM01
Microbiology 171, 001575 (2025); https://doi.org/10.1099/mic.0.001575
/content/journal/micro/10.1099/mic.0.001575
/content/journal/micro/10.1099/mic.0.001575
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