RESEARCH ARTICLE Natural Mutations in Streptococcus agalactiae Resulting in Abrogation of β Antigen Production

Streptococcus agalactiae genome encodes 21 two-component systems (TCS) and a varie-ty of regulatory proteins in order to control gene expression. One of the TCS, BgrRS, com-prising the BgrR DNA-binding regulatory protein and BgrS sensor histidine kinase, was discovered within a putative virulence island. BgrRS influences cell metabolism and posi-tively control the expression of bac gene, coding for β antigen at transcriptional level. Inacti-vation of bgrR abrogated bac gene expression and increased virulence properties of S. agalactiae. In this study, a total of 140 strains were screened for the presence of bac gene, and the TCS bgrR and bgrS genes. A total of 53 strains carried the bac, bgrR and bgrS genes. Most of them (48 strains) expressed β antigen, while five strains did not express β antigen. Three strains, in which bac gene sequence was intact, while bgrR and/or bgrS genes had mutations, and expression of β antigen was absent, were complemented with a constructed plasmid pBgrRS(P) encoding functionally active bgrR and bgrS gene alleles. This procedure restored expression of β antigen indicating the crucial regulatory role of TCS BgrRS. The complemented strain A49V/BgrRS demonstrated attenuated virulence i