title: Analyzing the modification of the Shewanella oneidensis MR-1 flagellar filament.
abstract: 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.
authors.affiliation: Department of Ecophysiology, Max Planck Institute for Terrestrial Microbiology, Marburg, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Bubendorfer
authors.forename: Mayumi
authors.initials: M
authors.lastname: Ishihara
authors.forename: Kim
authors.initials: K
authors.lastname: Dohlich
authors.forename: Christian
authors.initials: C
authors.lastname: Heiss
authors.forename: Jan
authors.initials: J
authors.lastname: Vogel
authors.forename: Federico
authors.initials: F
authors.lastname: Sastre
authors.forename: Maria
authors.initials: M
authors.lastname: Panico
authors.forename: Paul
authors.initials: P
authors.lastname: Hitchen
authors.forename: Anne
authors.initials: A
authors.lastname: Dell
authors.forename: Parastoo
authors.initials: P
authors.lastname: Azadi
authors.forename: Kai M
authors.initials: KM
authors.lastname: Thormann
mesh_terms: D000595:Amino Acid Sequence; D005407:Flagella / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005408:Flagellin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D005810:Multigene Family; D009154:Mutation; D011499:Protein Processing, Post-Translational; D020592:Shewanella / Q000166:cytology* / Q000235:genetics* / Q000378:metabolism
chemical_list: D005408:Flagellin
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