Autoaggregation inStreptococcus intermediusis driven by the Pel polysaccharide

Abstract

ABSTRACTThe Streptococcus Milleri Group (SMG) comprising ofStreptococcus intermedius, anginosusandconstellatusare commensal bacteria commonly found in healthy individuals. These bacteria are increasingly being recognized as opportunistic pathogens that can cause purulent infections at sterile body sites and have also been identified in the sputum of individuals with cystic fibrosis. Although the mechanisms of conversion to opportunistic pathogens are not well understood, auto-aggregation is a key driver of biofilm adhesion and cohesion in many Streptococci and Staphylococci. Here, we identify a gene cluster in theS. intermediusgenome with significant homology to thepeloperons inBacillus cereusandPseudomonas aeruginosa, which are required for Pel exopolysaccharide production and biofilm formation in these species. Characterization of a panel of clinicalS. intermediusstrains identified a range of aggregating phenotypes. Analysis of thepeloperon in the hyper-aggregating C1365 strain revealed that each of the canonicalpelDEADAFGgenes, but not the four additional genes are required for aggregation. Further, we demonstrate that C1365 produces a GalNAc-rich exopolysaccharide and that aggregates can be disrupted by the α1,4N-acetylgalactosaminidases, PelA and Sph3, but not other glycoside hydrolases, proteinase K or DNase I. Using an abscess model of mouse infection, we show that Pel driven aggregation leads to longer lasting infections, and that lack of Pel allows for the bacteria to be cleared more effectively. The polymer also affects how the bacteria interacts with the host immune system. Collectively, our data suggest that thepeloperon has relevancy toS. intermediuspathogenicity.