Abstract
Polar and lateral flagellin proteins from Aeromonas hydrophila strain AH-3 (serotype O34) were found to be glycosylated with different carbohydrate moieties. The lateral flagellin was modified at three sites in O-linkage, with a single monosaccharide of 376 Da, which we show to be a pseudaminic acid derivative. The polar flagellin was modified with a heterogeneous glycan, comprised of a heptasaccharide, linked through the same 376-Da sugar to the protein backbone, also in O-linkage. In-frame deletion mutants of pseudaminic acid biosynthetic genes pseB and pseF homologues resulted in abolition of polar and lateral flagellar formation by posttranscriptional regulation of the flagellins, which was restored by complementation with wild type pseB or F homologues or Campylobacter pseB and F.
Highlights
Aeromonas hydrophila flagellar glycosylation is crucial for flagellar production
We were able to demonstrate that both flagella, the constitutively expressed polar flagellum and the inducible lateral flagella, in A. hydrophila strain AH-3 showed O-glycosylations with two different glycans
The lateral flagella showed an O-glycosylation with a single monosaccharide of 376 Da linked in at least three different sites to the lateral flagellin, with one site of modification at serine 178
Summary
Results: Aeromonas hydrophila strain AH-3 lateral and polar flagellins are modified in O-linkage with a single monosaccharide or a heterogenous glycan (heptasaccharide), respectively. Significance: O-Glycosylation by two different glycans for polar and lateral flagellins and is crucial for flagellar biogenesis. Polar and lateral flagellin proteins from Aeromonas hydrophila strain AH-3 (serotype O34) were found to be glycosylated with different carbohydrate moieties. The lateral flagellin was modified at three sites in O-linkage, with a single monosaccharide of 376 Da, which we show to be a pseudaminic acid derivative. Inframe deletion mutants of pseudaminic acid biosynthetic genes pseB and pseF homologues resulted in abolition of polar and lateral flagellar formation by posttranscriptional regulation of the flagellins, which was restored by complementation with wild type pseB or F homologues or Campylobacter pseB and F
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