1. A general protein O- glycosylation machinery conserved in Burkholderia species improves bacterial fitness and elicits glycan immunogenicity in humans.
- Author
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Fathy Mohamed Y, Scott NE, Molinaro A, Creuzenet C, Ortega X, Lertmemongkolchai G, Tunney MM, Green H, Jones AM, DeShazer D, Currie BJ, Foster LJ, Ingram R, De Castro C, and Valvano MA
- Subjects
- Bacterial Proteins genetics, Chromatography, Liquid, Computational Biology, Glycoproteins genetics, Glycosylation, Humans, Mass Spectrometry, Mutation, Polysaccharides analysis, Recombinant Proteins genetics, Recombinant Proteins metabolism, Species Specificity, Bacterial Proteins metabolism, Burkholderia metabolism, Glycoproteins metabolism, Polysaccharides metabolism
- Abstract
The Burkholderia genus encompasses many Gram-negative bacteria living in the rhizosphere. Some Burkholderia species can cause life-threatening human infections, highlighting the need for clinical interventions targeting specific lipopolysaccharide proteins. Burkholderia cenocepacia O -linked protein glycosylation has been reported, but the chemical structure of the O -glycan and the machinery required for its biosynthesis are unknown and could reveal potential therapeutic targets. Here, using bioinformatics approaches, gene-knockout mutants, purified recombinant proteins, LC-MS-based analyses of O -glycans, and NMR-based structural analyses, we identified a B. cenocepacia O -glycosylation ( ogc ) gene cluster necessary for synthesis, assembly, and membrane translocation of a lipid-linked O -glycan, as well as its structure, which consists of a β-Gal-(1,3)-α-GalNAc-(1,3)-β-GalNAc trisaccharide. We demonstrate that the ogc cluster is conserved in the Burkholderia genus, and we confirm the production of glycoproteins with similar glycans in the Burkholderia species: B. thailandensis , B. gladioli , and B. pseudomallei Furthermore, we show that absence of protein O- glycosylation severely affects bacterial fitness and accelerates bacterial clearance in a Galleria mellonella larva infection model. Finally, our experiments revealed that patients infected with B. cenocepacia , Burkholderia multivorans , B. pseudomallei , or Burkholderia mallei develop O- glycan-specific antibodies. Together, these results highlight the importance of general protein O- glycosylation in the biology of the Burkholderia genus and its potential as a target for inhibition or immunotherapy approaches to control Burkholderia infections.
- Published
- 2019
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