Crystal structure of FliC flagellin from Pseudomonas aeruginosa and its implication in TLR5 binding and formation of the flagellar filament

Song, Wan Seok; Yoon, Sung‐il

doi:10.1016/j.bbrc.2014.01.008

Cited by 58 publications

(41 citation statements)

References 31 publications

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“…In contrast, FliD can be exchanged among a variety of bacteria without further problems (53). Finally, the detected direct interaction of the cytochrome c domain of NirS with both the proposed long N-and C-terminal helices of FliC might give insights into the structural interconnection of denitrification and motility (54). Overall, we assume that the periplasmic compartment represents a highly ordered and structured cellular organelle in Gram-negative bacteria.…”

Section: Discussionmentioning

confidence: 77%

A Periplasmic Complex of the Nitrite Reductase NirS, the Chaperone DnaK, and the Flagellum Protein FliC Is Essential for Flagellum Assembly and Motility in Pseudomonas aeruginosa

et al. 2015

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Pseudomonas aeruginosa is a ubiquitously occurring environmental bacterium and opportunistic pathogen responsible for various acute and chronic infections. Obviously, anaerobic energy generation via denitrification contributes to its ecological success. To investigate the structural basis for the interconnection of the denitrification machinery to other essential cellular processes, we have sought to identify the protein interaction partners of the denitrification enzyme nitrite reductase NirS in the periplasm. We employed NirS as an affinity-purifiable bait to identify interacting proteins in vivo. Results obtained revealed that both the flagellar structural protein FliC and the protein chaperone DnaK form a complex with NirS in the periplasm. The interacting domains of NirS and FliC were tentatively identified. The NirS-interacting stretch of amino acids lies within its cytochrome c domain. Motility assays and ultrastructure analyses revealed that a nirS mutant was defective in the formation of flagella and correspondingly in swimming motility. In contrast, the fliC mutant revealed an intact denitrification pathway. However, deletion of the nirF gene, coding for a heme d 1 biosynthetic enzyme, which leads to catalytically inactive NirS, did not abolish swimming ability. This pointed to a structural function for the NirS protein. FliC and NirS were found colocalized with DnaK at the cell surface of P. aeruginosa. A function of the detected periplasmic NirS-DnaK-FliC complex in flagellum formation and motility was concluded and discussed. IMPORTANCEPhysiological functions in Gram-negative bacteria are connected with the cellular compartment of the periplasm and its membranes. Central enzymatic steps of anaerobic energy generation and the motility mediated by flagellar activity use these cellular structures in addition to multiple other processes. Almost nothing is known about the protein network functionally connecting these processes in the periplasm. Here, we demonstrate the existence of a ternary complex consisting of the denitrifying enzyme NirS, the chaperone DnaK, and the flagellar protein FliC in the periplasm of the pathogenic bacterium P. aeruginosa. The dependence of flagellum formation and motility on the presence of an intact NirS was shown, structurally connecting both cellular processes, which are important for biofilm formation and pathogenicity of the bacterium. P seudomonas aeruginosa is a metabolically versatile bacterium inhabiting multiple environmental niches (1). It is known for its highly efficient growth in the absence of oxygen. Fast anaerobic growth is mediated via the utilization of different N-oxides as electron acceptors in the respiratory chains of denitrification (2). Anaerobic respiratory growth via denitrification also sustains biofilm formation on environmental surfaces, in the mucus in the lung of cystic fibrosis patients, on the epithelium of the urinary tract infected individuals, and in burn wounds (3).The periplasm, the cellular compartment bounded by the cytoplasmi...

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Section: Discussionmentioning

confidence: 77%

A Periplasmic Complex of the Nitrite Reductase NirS, the Chaperone DnaK, and the Flagellum Protein FliC Is Essential for Flagellum Assembly and Motility in Pseudomonas aeruginosa

et al. 2015

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“…The structures of several bacterial flagellins have been solved by X-ray crystallography (19)(20)(21), which involved removal of the N-and C-terminal D 0 domains. The structure of the complete Salmonella enterica serovar Typhimurium flagellin (i.e., also containing the D 0 domains) has also been solved by cryoelectron microscopy (22).…”

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confidence: 99%

A Critical Region in the FlaA Flagellin Facilitates Filament Formation of the Vibrio cholerae Flagellum

et al. 2018

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is a Gram-negative bacterium with a monotrichous flagellum that causes the human disease cholera. Flagellum-mediated motility is an integral part of the bacterial life cycle inside the host and in the aquatic environment. The flagellar filament is composed of five flagellin subunits (FlaA, FlaB, FlaC, FlaD, and FlaE); however, only FlaA is necessary and sufficient for filament synthesis. is transcribed from a class III flagellar promoter, whereas the other four flagellins are transcribed from class IV promoters. However, expressing from a class IV promoter still facilitated motility in a strain that was otherwise lacking all five flagellins (Δ). Furthermore, FlaA from (FlaA; 77% identity) supported motility of the Δ strain, whereas FlaA from (FlaA; 75% identity) did not, indicating that FlaA amino acid sequence is responsible for its critical role in flagellar synthesis. Chimeric proteins composed of different domains of FlaA and FlaD or FlaA revealed that the N-terminal D domain (D) contains an important region required for FlaA function. Further analyses of chimeric FlaA-FlaD proteins identified a lysine residue present at position 145 of the other flagellins but absent from FlaA that can prevent monofilament formation. Moreover, the D region of amino acids 87 to 153 of FlaA inserted into FlaA allows monofilament formation but not motility, apparently due to the lack of filament curvature. These results identify residues within the D domain that allow FlaA to fold into a functional filament structure and suggest that FlaA assists correct folding of the other flagellins. causes the severe diarrheal disease cholera. Its ability to swim is mediated by rotation of a polar flagellum, and this motility is integral to its ability to cause disease and persist in the environment. The current studies illuminate how one specific flagellin (FlaA) within a multiflagellin structure mediates formation of the flagellar filament, thus allowing to swim. This knowledge can lead to safer vaccines and potential therapeutics to inhibit cholera.

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“…36 However, the structure of three Tpflagellins was distinguished in residues 127-198 where TpFlaB1 contained two β-hairpin ( Figure 1B Tpflagellins were structurally similar to its orthologs (Stflagellin, Paflagellin, Ssflagellin, Sdflagellin and Bsflagellin) in that they contained three α-helices bundle structure ( Figure 1B,C). [36][37][38][48][49][50] The…”

Section: Modelling Of Tpflagellins Structures and Structural Comparmentioning

confidence: 99%

The N‐terminal D1 domain of Treponema pallidum flagellin binding to TLR5 is required but not sufficient in activation of TLR5

Xie

Tan

et al. 2019

J Cellular Molecular Medi

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Syphilis is a chronic bacterial infection caused by Treponema pallidum (T pallidum) and the pathogenesis that T pallidum infection induces immunopathological damages in skin and other tissues remains unclear. We have previously reported that recombinant flagellins of T pallidum can elicit IL‐6 and IL‐8 transcriptions via TLR5 pathway. To identify the domains which induced the pro‐inflammatory activity and the importance of the interactions between TLR5 and domains, homology‐based modelling and comparative structural analyses revealed that Tpflagellins can combine with TLR5 directly. Deletion mutations showed that the ND1 domain binding to TLR5 is required but not sufficient in TLR5 activation. Moreover, site‐directed mutagenesis analysis indicated that the arginine residue (Tpflagellins R89) of the ND1 domain and its adjacent residues (Tpflagellins L93 and E113) constitute a hot spot that elicits IL‐6, IL‐8 transcriptions and TLR5 activation, and affects the binding of Tpflagellins to TLR5. Taken together, these results give insight into the pathogenesis of T pallidum and may contribute to the future design of Tpflagellins‐based therapeutics and syphilis vaccine.

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Crystal structure of FliC flagellin from Pseudomonas aeruginosa and its implication in TLR5 binding and formation of the flagellar filament

Cited by 58 publications

References 31 publications

A Periplasmic Complex of the Nitrite Reductase NirS, the Chaperone DnaK, and the Flagellum Protein FliC Is Essential for Flagellum Assembly and Motility in Pseudomonas aeruginosa

A Periplasmic Complex of the Nitrite Reductase NirS, the Chaperone DnaK, and the Flagellum Protein FliC Is Essential for Flagellum Assembly and Motility in Pseudomonas aeruginosa

A Critical Region in the FlaA Flagellin Facilitates Filament Formation of the Vibrio cholerae Flagellum

The N‐terminal D1 domain of Treponema pallidum flagellin binding to TLR5 is required but not sufficient in activation of TLR5

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