Regulation of the ß-hemolysin gene cluster of Streptococcus anginosus by CcpA.

Streptococcus anginosus is increasingly recognized as an opportunistic pathogen. However, our knowledge about virulence determinants in this species is scarce. One exception is the streptolysin-S (SLS) homologue responsible for the ß-hemolytic phenotype of the S. anginosus type strain. In S. anginosus the expression of the hemolysin is reduced in the presence of high glucose concentrations. To investigate the genetic mechanism of the hemolysin repression we created an isogenic ccpA deletion strain. In contrast to the wild type strain, this mutant exhibits hemolytic activity in presence of up to 25 mM glucose supplementation, a phenotype that could be reverted by ccpA complementation. To further demonstrate that CcpA directly regulates the hemolysin expression, we performed an in silico analysis of the promoter of the SLS gene cluster and we verified the binding of CcpA to the promoter by electrophoretic mobility shift assays. This allowed us to define the CcpA binding site in the SLS promoter region of S. anginosus. In conclusion, we report for the first time the characterization of a potential virulence regulator in S. anginosus.