title: Gonococcal rfaF mutants express Rd2 chemotype LPS and do not enter epithelial host cells.
abstract: We have investigated the function of the Isi-1 gene of Neisseria gonorrhoeae previously implicated in lipopolysaccharide (LPS)-inner-core biosynthesis (Petricoin et al., 1991). Disruption of the gene in gonococcal strain MS11 resulted in the production of LPS that migrated faster than that from an isogenic galE mutant, typical for a mutation that influences the inner-core region. Complementation of a panel of Salmonella typhimurium mutants with defined defects in rfa loci demonstrated conclusively that the Isi-1 gene of MS11 is functionally homologous to the rfaF gene, which encodes heptosyltransferase II in both E. coli and S. typhimurium. Comparison of deduced amino acid sequences of the gonococcal and the Salmonella RfaF demonstrated 70% similarity, including 47% identical amino acid residues. Immunochemical analysis of the LPS using monoclonal antibodies directed against chemically defined inner-core glycoconjugates revealed that the gonococcal and Salmonella Rd2-chemotypes were antigenically similar, further extending the genetic and functional homology. Infection experiments in vitro demonstrated that the Isi-1 mutant could not invade human Chang epithelial cells despite expression of a genetically defined invasion-promoting gonococcal opacity protein. These data imply that the LPS phenotype is a critical factor for gonococcal invasiveness.
authors.affiliation: Max-Planck-Institut für Biologie, Abteilung Infektionsbiologie, Tübingen, Germany.
authors.forename: E T
authors.initials: ET
authors.lastname: Schwan
authors.forename: B D
authors.initials: BD
authors.lastname: Robertson
authors.forename: H
authors.initials: H
authors.lastname: Brade
authors.forename: J P
authors.initials: JP
authors.lastname: van Putten
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D016695:Glycosyltransferases / Q000235:genetics / Q000502:physiology; D008070:Lipopolysaccharides / Q000096:biosynthesis* / Q000145:classification / Q000276:immunology; D008969:Molecular Sequence Data; D009344:Neisseria gonorrhoeae / Q000145:classification / Q000235:genetics* / Q000276:immunology / Q000472:pathogenicity; D012486:Salmonella typhimurium / Q000235:genetics / Q000276:immunology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014774:Virulence / Q000235:genetics
chemical_list: D008070:Lipopolysaccharides; D016695:Glycosyltransferases; C086735:heptosyltransferase