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bioRxiv · 10.1101/2022.03.25.485807

Genetic glyco-profiling and rewiring of insulated flagellin glycosylation pathways

Kint, N.·Dubois, T.·Viollier, P. H.
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Abstract

Glycosylation of surface structures diversifies cells chemically and physically. Sialic acids commonly serve as glycosyl donors, particularly pseudaminic (Pse) or legionaminic acid (Leg) that prominently decorate eubacterial and archaeal surface layers or appendages. We investigated a new class of FlmG protein glycosyltransferases that modify flagellin, the structural subunit of the flagellar filament. Functional insulation of orthologous Pse and Leg biosynthesis pathways accounted for the flagellin glycosylation specificity and motility conferred by the cognate FlmG in the -proteobacteria Caulobacter crescentus and Brevundimonas subvibrioides, respectively. Exploiting these functions, we conducted genetic glyco-profiling to classify Pse or Leg biosynthesis pathways and we used heterologous reconstitution experiments to unearth a signature determinant of Leg biosynthesis in eubacteria and archaea. These findings and our chimeric FlmG analyses reveal two modular determinants that govern flagellin glycosyltransferase specificity: a glycosyltransferase domain that accepts either Leg or Pse and that uses specialized flagellin-binding domain to identify the substrate.

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Kint, N., Dubois, T., Viollier, P. H.. 2022-03-26. Genetic glyco-profiling and rewiring of insulated flagellin glycosylation pathways. https://doi.org/10.1101/2022.03.25.485807

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