Carbohydrate engineering of the recognition motifs in streptococcal co-aggregation receptor polysaccharides.
The cell wall polysaccharides of certain oral streptococci function as receptors for the lectin-like surface adhesins on other members of the oral biofilm community. Recognition of these receptor polysaccharides (RPS) depends on the presence of a host-like motif, either GalNAcbeta1-3Gal (Gn) or Galbeta1-3GalNAc (G), within the oligosaccharide repeating units of different RPS structural types. Type 2Gn RPS of Streptococcus gordonii 38 and type 2G RPS of Streptococcus oralis J22 are composed of heptasaccharide repeats that are identical except for their host-like motifs. In the current investigation, the genes for the glycosyltransferases that synthesize these motifs were identified by high-resolution nuclear magnetic resonance (NMR) analysis of genetically altered polysaccharides. RPS production was switched from type 2Gn to 2G by replacing wefC and wefD in the type 2Gn gene cluster of S. gordonii 38 with wefF and wefG from the type 2G cluster of S. oralis J22. Disruption of either wefC or wefF abolished cell surface RPS production. In contrast, disruption of wefD in the type 2Gn cluster or wefG in the type 2G cluster eliminated beta-GalNAc from the Gn motif or beta-Gal from the G motif, resulting in mutant polysaccharides with hexa- rather than heptasaccharide subunits. The mutant polysaccharides reacted like wild-type RPS with rabbit antibodies against type 2Gn or 2G RPS but were inactive as co-aggregation receptors. Additional mutant polysaccharides with GalNAcbeta1-3GalNAc or Galbeta1-3Gal recognition motifs were engineered by replacing wefC in the type 2Gn cluster with wefF or wefF in the type 2G cluster with wefC respectively. The reactions of these genetically modified polysaccharides as antigens and receptors provide further insight into the structural basis of RPS function.
Yoshida
Yasuo
Y
Oral Infection and Immunity Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892, USA.
Ganguly
Soumya
S
Bush
C Allen
CA
Cisar
John O
JO
D020816:Amino Acid Motifs; D001422:Bacterial Adhesion / Q000235:genetics; D002240:Carbohydrate Sequence; D002473:Cell Wall / Q000737:chemistry; D004269:DNA, Bacterial; D017353:Gene Deletion; D005798:Genes, Bacterial; D005818:Genetic Engineering; D016695:Glycosyltransferases / Q000235:genetics; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D011135:Polysaccharides, Bacterial / Q000737:chemistry / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000235:genetics; D011995:Recombination, Genetic; D017422:Sequence Analysis, DNA; D013291:Streptococcus / Q000235:genetics* / Q000378:metabolism*
D004269:DNA, Bacterial; D011135:Polysaccharides, Bacterial; D011956:Receptors, Cell Surface; D016695:Glycosyltransferases