title: Maf-dependent bacterial flagellin glycosylation occurs before chaperone binding and flagellar T3SS export.
abstract: Bacterial swimming is mediated by rotation of a filament that is assembled via polymerization of flagellin monomers after secretion via a dedicated flagellar Type III secretion system. Several bacteria decorate their flagellin with sialic acid related sugars that is essential for motility. Aeromonas caviae is a model organism for this process as it contains a genetically simple glycosylation system and decorates its flagellin with pseudaminic acid (Pse). The link between flagellin glycosylation and export has yet to be fully determined. We examined the role of glycosylation in the export and assembly process in a strain lacking Maf1, a protein involved in the transfer of Pse onto flagellin at the later stages of the glycosylation pathway. Immunoblotting, established that glycosylation is not required for flagellin export but is essential for filament assembly since non-glycosylated flagellin is still secreted. Maf1 interacts directly with its flagellin substrate in vivo, even in the absence of pseudaminic acid. Flagellin glycosylation in a flagellin chaperone mutant (flaJ) indicated that glycosylation occurs in the cytoplasm before chaperone binding and protein secretion. Preferential chaperone binding to glycosylated flagellin revealed its crucial role, indicating that this system has evolved to favour secretion of the polymerization competent glycosylated form.
authors: Department of Infection and Immunity, University of Sheffield, Sheffield, S10 2RX, UK.
authors: Jennifer L
authors: JL
authors: Parker
authors: Rebecca C
authors: RC
authors: Lowry
authors: Narciso A S
authors: NA
authors: Couto
authors: Phillip C
authors: PC
authors: Wright
authors: Graham P
authors: GP
authors: Stafford
authors: Jonathan G
authors: JG
authors: Shaw
mesh_terms: D057585:Aeromonas caviae / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D058947:Bacterial Secretion Systems; D005408:Flagellin / Q000378:metabolism*; D017353:Gene Deletion; D006031:Glycosylation; D011485:Protein Binding; D025941:Protein Interaction Mapping; D055503:Protein Multimerization; D021381:Protein Transport; D013400:Sugar Acids / Q000378:metabolism
chemical_list: C513444:5,7-diacetamido-3,5,7,9-tetradeoxynonulosonic acid; D001426:Bacterial Proteins; D058947:Bacterial Secretion Systems; D013400:Sugar Acids; D005408:Flagellin