Streptococcus agalactiae capsule polymer length and attachment is determined by the proteins CpsABCD [Glycobiology and Extracellular Matrices]

February 9th, 2015 by Toniolo, C., Balducci, E., Romano, M. R., Proietti, D., Ferlenghi, I., Grandi, G., Berti, F., Margarit Y.Ros, I., Janulczyk, R.

The production of capsular polysaccharides (CPS) or secreted exopolysaccharides is ubiquitous in bacteria, and the Wzy pathway constitutes a prototypical mechanism to produce these structures. Despite the differences in polysaccharide composition among species, a group of proteins involved in this pathway is well conserved. Streptococcus agalactiae (GBS) produces a CPS which represents the main virulence factor of the bacterium and is a prime target in current vaccine development. We used this human pathogen to investigate the roles and potential interdependencies of the conserved proteins CpsABCD encoded in the cps operon, by developing knock out and functional mutant strains. The mutant strains were examined for CPS quantity, size, and attachment to the cell surface, as well as CpsD phosphorylation. We observed that CpsB, C and D compose a phosphoregulatory system where the CpsD autokinase phosphorylates its C-terminal tyrosines in a CpsC-dependent manner. These Tyr residues are also the target of the cognate CpsB phosphatase. An interaction between CpsD and CpsC was observed, and the phosphorylation state of CpsD influenced the subsequent action of CpsC. The CpsC extracellular domain appeared necessary for the production of high molecular weight polysaccharides by influencing CpsA-mediated attachment of the CPS to the bacterial cell surface. In conclusion, while having no impact on cps transcription or the synthesis of the basal repeating unit, we suggest that these proteins are fine-tuning the last steps of CPS biosynthesis, i.e. the balance between polymerization and attachment to the cell wall.
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