A serum-sensitive, sialyltransferase-deficient mutant of Neisseria gonorrhoeae defective in conversion to serum resistance by CMP-NANA or blood cell extracts.
A stable, sialyltransferase-deficient mutant of Neisseria gonorrhoeae strain F62 totally defective in CMP-NANA-dependent lipopolysaccharide (LPS) sialylation was isolated by insertion mutagenesis with transposon Tn 1545-delta 3 and screened for unlabelled colonies following incubation with CMP-14C-NANA. In contrast to the parental strain which became serum resistant on incubation with CMP-NANA or blood cell extracts, the mutant, JB1, remained serum sensitive. French press extracts of strain F62 catalysed LPS sialylation, but corresponding extracts of the mutant were inactive. Five LPS components were detected by SDS-PAGE in the parental strain. Five components of the same Mr were also found in the mutant. Three identical components were detected by Western blotting using MAb 3F11, which recognizes the Gal beta 1-4GlcNAc groups in the conserved LPS components of F62 which can be sialylated. The mutant, JB1, is therefore deficient in the sialyltransferase that is essential for both LPS sialylation and conversion of serum-sensitive gonococci to serum resistance by either CMP-NANA or blood cell extracts. No evidence was obtained for an LPS sialylation pathway by blood cell extracts that is independent of CMP-NANA.
Bramley
J
J
School of Biochemistry, University of Birmingham, U.K.
Demarco de Hormaeche
R
R
Constantinidou
C
C
Nassif
X
X
Parsons
N
N
Jones
P
P
Smith
H
H
Cole
J
J
D000911:Antibodies, Monoclonal; D001770:Blood Bactericidal Activity; D002240:Carbohydrate Sequence; D003569:Cytidine Monophosphate N-Acetylneuraminic Acid / Q000494:pharmacology; D004251:DNA Transposable Elements; D006801:Humans; D066298:In Vitro Techniques; D008070:Lipopolysaccharides / Q000737:chemistry; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009344:Neisseria gonorrhoeae / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D012794:Sialic Acids / Q000737:chemistry; D012799:Sialyltransferases / Q000235:genetics
D000911:Antibodies, Monoclonal; D004251:DNA Transposable Elements; D008070:Lipopolysaccharides; D012794:Sialic Acids; D003569:Cytidine Monophosphate N-Acetylneuraminic Acid; D012799:Sialyltransferases