Identification of novel glycosyltransferases required for assembly of the Pasteurella multocida A:1 lipopolysaccharide and their involvement in virulence.
We previously determined the structure of the Pasteurella multocida Heddleston type 1 lipopolysaccharide (LPS) molecule and characterized some of the transferases essential for LPS biosynthesis. We also showed that P. multocida strains expressing truncated LPS display reduced virulence. Here, we have identified all of the remaining glycosyltransferases required for synthesis of the oligosaccharide extension of the P. multocida Heddleston type 1 LPS, including a novel alpha-1,6 glucosyltransferase, a beta-1,4 glucosyltransferase, a putative bifunctional galactosyltransferase, and two heptosyltransferases. In addition, we identified a novel oligosaccharide extension expressed only in a heptosyltransferase (hptE) mutant background. All of the analyzed mutants expressing LPS with a truncated main oligosaccharide extension displayed reduced virulence, but those expressing LPS with an intact heptose side chain were able to persist for long periods in muscle tissue. The hptC mutant, which expressed LPS with the shortest oligosaccharide extension and no heptose side chain, was unable to persist on the muscle or cause any disease. Furthermore, all of the mutants displayed increased sensitivity to the chicken antimicrobial peptide fowlicidin 1, with mutants expressing highly truncated LPS being the most sensitive.
Boyce
John D
JD
Australian Research Council Centre of Excellence in Structural and Functional Microbial Genomics, Monash University, VIC 3800, Australia.
Harper
Marina
M
St Michael
Frank
F
John
Marietta
M
Aubry
Annie
A
Parnas
Henrietta
H
Logan
Susan M
SM
Wilkie
Ian W
IW
Ford
Mark
M
Cox
Andrew D
AD
Adler
Ben
B
D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002645:Chickens; D005700:Galactosyltransferases / Q000235:genetics / Q000378:metabolism; D005964:Glucosyltransferases / Q000235:genetics / Q000378:metabolism; D016695:Glycosyltransferases / Q000235:genetics / Q000378:metabolism; D008070:Lipopolysaccharides / Q000096:biosynthesis* / Q000737:chemistry; D008826:Microbial Sensitivity Tests; D009154:Mutation; D010326:Pasteurella Infections / Q000382:microbiology / Q000503:physiopathology / Q000662:veterinary*; D016979:Pasteurella multocida / Q000201:enzymology / Q000235:genetics / Q000472:pathogenicity*; D011201:Poultry Diseases / Q000382:microbiology / Q000503:physiopathology*; D014774:Virulence
D023181:Antimicrobial Cationic Peptides; D001426:Bacterial Proteins; D008070:Lipopolysaccharides; D016695:Glycosyltransferases; C036655:1,3-alpha-D-glucan synthase; D005700:Galactosyltransferases; D005964:Glucosyltransferases; C086735:heptosyltransferase