Differential Glycosylation of Polar and Lateral Flagellins in Aeromonas hydrophila AH-3

Wilhelms, Markus; Fulton, Kelly M.; Twine, Susan M.; Tomás, Juan M.; Merino, Susana

doi:10.1074/jbc.m112.376525

Cited by 36 publications

(74 citation statements)

References 37 publications

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“…For this organism, pseudaminic acid (Pse) is among the predominant O-linked glycans, and the linkage occurs prior to the transport of the flagellin and assembly of the filament. The flagellin of Helicobacter pylori has been demonstrated to be similarly decorated with Pse [17], [19]–[21], and a recent study on A. hydrophila also strongly suggests a role of Pse in flagellin glycosylation of both polar and lateral flagellar systems [22].…”

Section: Introductionmentioning

confidence: 95%

Analyzing the Modification of the Shewanella oneidensis MR-1 Flagellar Filament

et al. 2013

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The unsheathed flagellar filament of Shewanella oneidensis MR-1 is composed of two highly homologous flagellins, FlaA, and the major structural unit, FlaB. We identified a gene cluster, SO_3261-SO_3265 (now sfmABCDE), that is required for the formation of a fully functional filament and for motility. The predicted function of the corresponding gene products strongly indicated a role in flagellin modification. Accordingly, loss of sfmABCDE results in a significant mass shift of both FlaA and FlaB. Mass spectroscopy analysis and single residue substitutions identified five serine residues in both flagellins that are modified via O-linkage. Modeling of the flagellin structures strongly suggests that at least four of the modified residues are exposed to the filament’s surface. However, none of the five serine residues solely is crucial for function and assembly. Structural analysis of the flagellin modification revealed that it likely contains a nonulosonic acid (274 Da) linked to each glycosylated serine. The putative nonulosonic acid is further substituted with a 236 Da moiety which can carry additional methyl groups (250 Da, 264 Da). In addition, at least 5 lysine residues in FlaB and one in FlaA were found to be methylated. Based on homology comparisons we suggest that smfABCDE is required for species-specific flagellin modification in S. oneidensis MR-1.

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Section: Introductionmentioning

confidence: 95%

Analyzing the Modification of the Shewanella oneidensis MR-1 Flagellar Filament

et al. 2013

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“…Since then, Pse and derivatives have been frequently found as constituents of cell surface glycoconjugates such as pili (Horzempa et al 2006), flagella (Thibault et al 2001; Schirm et al 2003), LPS (Knirel et al 2003) and capsular polysaccharides (Kiss et al 2001; Kenyon et al 2014). In Campylobacter jejuni, Helicobacter pylori and Aeromonas spp., heavy glycosylation of the flagellin protein with Pse has been shown to be an essential requirement for flagellar assembly and, thus, motility and virulence of these pathogens (Guerry et al 2006; Wilhelms et al 2012; Lowry et al 2014). …”

Section: Introductionmentioning

confidence: 99%

Tannerella forsythiastrains display different cell-surface nonulosonic acids: biosynthetic pathway characterization and first insight into biological implications

et al. 2016

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Tannerella forsythia is an anaerobic, Gram-negative periodontal pathogen. A unique O-linked oligosaccharide decorates the bacterium’s cell surface proteins and was shown to modulate the host immune response. In our study, we investigated the biosynthesis of the nonulosonic acid (NulO) present at the terminal position of this glycan. A bioinformatic analysis of T. forsythia genomes revealed a gene locus for the synthesis of pseudaminic acid (Pse) in the type strain ATCC 43037 while strains FDC 92A2 and UB4 possess a locus for the synthesis of legionaminic acid (Leg) instead. In contrast to the NulO in ATCC 43037, which has been previously identified as a Pse derivative (5-N-acetimidoyl-7-N-glyceroyl-3,5,7,9-tetradeoxy-l-glycero-l-manno-NulO), glycan analysis of strain UB4 performed in this study indicated a 350-Da, possibly N-glycolyl Leg (3,5,7,9-tetradeoxy-d-glycero-d-galacto-NulO) derivative with unknown C5,7 N-acyl moieties. We have expressed, purified and characterized enzymes of both NulO pathways to confirm these genes’ functions. Using capillary electrophoresis (CE), CE–mass spectrometry and NMR spectroscopy, our studies revealed that Pse biosynthesis in ATCC 43037 essentially follows the UDP-sugar route described in Helicobacter pylori, while the pathway in strain FDC 92A2 corresponds to Leg biosynthesis in Campylobacter jejuni involving GDP-sugar intermediates. To demonstrate that the NulO biosynthesis enzymes are functional in vivo, we created knockout mutants resulting in glycans lacking the respective NulO. Compared to the wild-type strains, the mutants exhibited significantly reduced biofilm formation on mucin-coated surfaces, suggestive of their involvement in host-pathogen interactions or host survival. This study contributes to understanding possible biological roles of bacterial NulOs.

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“…In contrast to the glycosylated polar flagellin of A. caviae, the polar flagellin of A. hydrophila AH-3 appears to be more complicated as they are modified with a heterogeneous glycan that consists of a heptasaccharide linked to a pseudaminic acid derivative of 376 Da (Wilhelms, Fulton, Twine, Tomás, & Merino, 2012). In addition the flagellin glycosylation island of A. hydrophila, AH-3 is more complex with the presence of several extra genes that could possibly account for the heterogeneous glycan or the modified pseudaminic acid derivative, as well as the presence of two maf gene homologues that may be required for transferring specific sugars on to the flagellin monomers .…”

Section: Flagellin Glycosylationmentioning

confidence: 94%

“…More recently, findings have shown that this aberrant migration is due to the modification of lateral flagellin of A. hydrophila via O-linked glycosylation on to serine and threonine residues. A. hydrophila lateral flagellins are glycosylated exclusively with a 376 Da derivative of pseudaminic acid that is also used to modify the polar flagellin (Wilhelms et al, 2012), although it is not known whether the two flagella systems (polar and lateral), share glycosylation machinery. It appears that glycosylation of the lateral flagellin in this strain of A. hydrophila is essential for swarming motility over surfaces.…”

Section: Lateral Flagellamentioning

confidence: 99%