Molecular analysis of a region of the group B streptococcus chromosome involved in type III capsule expression

Abstract

Type III group B streptococci (GBS) are the most common cause of neonatal sepsis and meningitis in the United States. The important role of the type III polysaccharide capsule and of the terminal sialic acid moiety of the capsule in the virulence of GBS has been demonstrated by using Tn916 mutagenesis. Several of the transposon insertion sites that resulted in defective type III capsule synthesis were located in a 30-kilobase (kb) region of the chromosome. Hybridization analysis of two other type III strains that differed in their relative virulence and of GBS serotypes Ia, Ib, Ic, and II showed that this region of the chromosome was highly conserved. A repetitive 1.4-kb sequence was found only in the 30-kb region of the more virulent type III strain, COH 1. The Escherichia coli maxicell in vivo expression system and an in vitro coupled transcription-translation system successfully identified the proteins expressed from the 30-kb region. Comparison of the proteins expressed from the same DNA fragments in these two assays indicated that some of these proteins may contain leader sequences that would ultimately result in their secretion to the cell surface. Identification and further characterization of the genes and their products will provide the foundation for understanding the genetic and biochemical events in GBS capsular polysaccharide production.