Sialylation of lipooligosaccharide cores affects immunogenicity and serum resistance of Campylobacter jejuni.
Three genes involved in biosynthesis of the lipooligosaccharide (LOS) core of Campylobacter jejuni MSC57360, the type strain of the HS:1 serotype, whose structure mimics GM(2) ganglioside, have been cloned and characterized. Mutation of genes encoding proteins with homology to a sialyl transferase (cstII) and a putative N-acetylmannosamine synthetase (neuC1), part of the biosynthetic pathway of N-acetylneuraminic acid (NeuNAc), have identical phenotypes. The LOS cores of these mutants display identical changes in electrophoretic mobility, loss of reactivity with cholera toxin (CT), and enhanced immunoreactivity with a hyperimmune polyclonal antiserum generated against whole cells of C. jejuni MSC57360. Loss of sialic acid in the core of the neuC1 mutant was confirmed by fast atom bombardment mass spectrometry. Mutation of a gene encoding a putative beta-1,4-N-acetylgalactosaminyltransferase (Cgt) resulted in LOS cores intermediate in electrophoretic mobility between that of wild type and the mutants lacking NeuNAc, loss of reactivity with CT, and a reduced immunoreactivity with hyperimmune antiserum. Chemical analyses confirmed the loss of N-acetylgalactosamine (GalNAc) and the presence of NeuNAc in the cgt mutant. These data suggest that the Cgt enzyme is capable of transferring GalNAc to an acceptor with or without NeuNAc and that the Cst enzyme is capable of transferring NeuNAc to an acceptor with or without GalNAc. A mutant with a nonsialylated LOS core is more sensitive to the bactericidal effects of human sera than the wild type or the mutant lacking GalNAc.
Guerry
P
P
Enteric Diseases Department, Naval Medical Research Center, Silver Spring, Maryland 20910, USA. guerryp@nmrc.navy.mil
Ewing
C P
CP
Hickey
T E
TE
Prendergast
M M
MM
Moran
A P
AP
D000818:Animals; D001770:Blood Bactericidal Activity; D016123:Campylobacter jejuni / Q000276:immunology* / Q000472:pathogenicity; D005408:Flagellin / Q000235:genetics; D008070:Lipopolysaccharides / Q000737:chemistry*; D016254:Mutagenesis, Insertional; D019158:N-Acetylneuraminic Acid / Q000378:metabolism*; D016366:Open Reading Frames; D011817:Rabbits
D008070:Lipopolysaccharides; C023023:lipid-linked oligosaccharides; D005408:Flagellin; D019158:N-Acetylneuraminic Acid