Streptococcus constellatus is a member of the Anginosus group streptococci (AGS) and primarily inhabits the human oral cavity. S. constellatus is composed of three subspecies: S. constellatus subsp. constellatus (SCC), S. constellatus subsp. pharyngis and the newly described subspecies S. constellatus subsp. viborgensis. Although previous studies have established that SCC contains b-haemolytic strains, the factor(s) responsible for b-haemolysis in b-haemolytic SCC (b-SCC) has yet to be clarified. Recently, we discovered that a streptolysin S (SLS) homologue is the b-haemolytic factor of b-haemolytic Streptococcus anginosus subsp. anginosus (b-SAA), another member of the AGS. Furthermore, because previous studies have suggested that other AGS species, except for Streptococcus intermedius, do not possess a haemolysin(s) belonging to the family of cholesterol-dependent cytolysins, we hypothesized that, as with b-SAA, the SLS homologue is the b-haemolytic factor of b-SCC, and therefore aimed to investigate and characterize the haemolytic factor of b-SCC in the present study. PCR amplification revealed that all of the tested b-SCC strains were positive for the sagA homologue of SCC (sagA SCC ). Further investigations using b-SCC strain W277 were conducted to elucidate the relationship between sagA SCC and b-haemolysis by constructing sagA SCC deletion mutants, which completely lost bhaemolytic activity. This loss of b-haemolytic activity was restored by trans-complementation of sagA SCC . Furthermore, a co-cultivation assay established that the cytotoxicity of b-SCC was clearly dependent on the presence of sagA SCC . These results demonstrate that sagA SCC is the factor responsible for b-SCC b-haemolysis and cytotoxicity.
INTRODUCTIONAnginosus group streptococci (AGS) are commensal bacteria in the human oral cavity. Current classification assigns AGS to three species, including five subspecies (Jensen et al., 2013): Streptococcus anginosus subsp. anginosus (SAA), S. anginosus subsp. whileyi (SAW), Streptococcus constellatus Abbreviations: AGS, Anginosus group streptococci; CSP, competence-stimulating peptide; erm, erythromycin resistance gene cassette; GAS, group A streptococci; GBS, group B streptococci; PGS, Pyogenic group streptococci; SAA, Streptococcus anginosus subsp. anginosus; SAW, Streptococcus anginosus subsp. whileyi; SCC, Streptococcus constellatus subsp. constellatus; SCP, Streptococcus constellatus subsp. pharyngis; SCV, Streptococcus constellatus subsp. viborgensis; SLS, streptolysin S.The GenBank/EMBL/DDBJ accession numbers for the sag operon homologue from b-haemolytic SCC strain W277, the sagA homologues from bhaemolytic SCC strains HW74 and 1340, and the sag operon homologues from the SCP type strain MM9889a T (5SK1060 T ), the SAW strain MAS624 and the SCV type strain SK1359 T are AB389362, AB389363, AB389364, AB389365, AB389366 and AB860419, respectively. (2014), 160, 980-991
Microbiology