The wild-type flagellar filament of the Firmicute Kurthia at 2.8 Å resolution in vivo

Blum, Thorsten B.; Filippidou, Sevasti; Fatton, Mathilda; Junier, Pilar; Abrahams, Jan Pieter

doi:10.1038/s41598-019-51440-1

Cited by 16 publications

(25 citation statements)

References 34 publications

(35 reference statements)

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“…S3B). Therefore, initial estimates for helical symmetry were taken from homologous flagellins of known structure (9,(54)(55)(56)). An initial model was obtained using (Fig.…”

Section: Data Collection and Processingmentioning

confidence: 99%

“…A handful of 11-start flagellin structures have been determined, including flagellar filaments from B. subtilis, S. enterica, Pseudomonas aeruginosa, and Kurthia sp. strain 11kri321 (9,(54)(55)(56). In the case of the B. subtilis, S. enterica, and P. aeruginosa reconstructions, single residue substitutions were used to straighten the filaments by locking the 11-start protofilament into either right (R-type) or left (L-type) handedness conformations, which yielded a uniform filament onto which helical symmetry could be more effectively determined and applied during structure determination (55,56).…”

Section: Introductionmentioning

confidence: 99%

“…Recently, a high-resolution reconstruction of the flagellar filament from Kurthia sp. found the corresponding 11-start protofilament to be nearly parallel to the helical axis with a rotation of approximately +0.5°, which could be considered an outlier from all other successful helical reconstructions to date(9,(54)(55)(56).…”

mentioning

confidence: 98%

“…These small differences in helical symmetry, however, can exert a large effect in the handedness of the 11-start protofilament. Until recently, all determined structures of 11-start flagellins fell into two classes: left-handed structures with an 11-start rotation of approximately -1.7° or right -handed structures with an 11-start rotation of approximately +3.8°(9,(54)(55)(56). These two dominant classes of 11-start protofilaments strongly influenced the field's view of filament structure, where toggling between two defined states was thought to play a functional role in bacterial motility(2).…”

mentioning

confidence: 99%

“…Subsequent to auto-refinement and per-particle CTF and motion correction, another round of 3-D classification without alignment was used to further curate the dataset, prior to re-extraction to the native pixel size of 0.87 Å and a final round of masked refinement and post-processing. Subsequent to reconstruction, a final round of 2-D classificationwithout alignment was used to ensure the refined particle stack exhibited 2-D classes with amplitude spectra that were qualitatively similar to amplitude spectra obtained from naïve 2-D classification before imposing any estimates of helical symmetry from homologous structures(9,(54)(55)(56)…”

mentioning

confidence: 99%

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Flagellar structures from the bacteriumCaulobacter crescentusand implications for phage ϕCbK predation of multi-flagellin bacteria

Eric

Nicoleta

Sanchez

et al. 2020

Preprint

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Caulobacter crescentus is a gram-negative alpha-proteobacterium that commonly lives in oligotrophic fresh and salt-water environments. C. crescentus is a host to many bacteriophages, including ϕCbK and ϕCbK-like bacteriophages, which first adsorb to cells by interaction with the bacterial flagellum. It is commonly thought that the six paralogs of the flagellin gene present in C. crescentus are important for bacteriophage evasion. Here, we show that deletion of specific flagellins in C. crescentus can indeed attenuate ϕCbK adsorption efficiency, although no single deletion completely ablates ϕCbK adsorption. Thus, bacteriophage ϕCbK likely recognizes a common motif amongst the six known flagellins in C. crescentus with varying degrees of efficiency. Interestingly, we observe that most deletion strains still generate flagellar filaments, with the exception of a strain that contains only the most divergent flagellin, FljJ, or a strain that contains only FljN and FljO. To visualize the surface residues that are likely recognized by ϕCbK, we determined two high-resolution structures of the FljK filament, with and without an amino acid substitution that induces straightening of the filament. We observe post-translational modifications on conserved surface threonine residues of FljK that are likely O-linked glycans. The possibility of interplay between these modifications and ϕCbK adsorption is discussed. We also determined the structure of a filament composed of a heterogeneous mixture of FljK and FljL, the final resolution of which was limited to approximately 4.6 Å. Altogether, this work builds a platform for future investigation of how phage ϕCbK infects C. crescentus at the molecular level.

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“…S3B). Therefore, initial estimates for helical symmetry were taken from homologous flagellins of known structure (9,(54)(55)(56)). An initial model was obtained using (Fig.…”

Section: Data Collection and Processingmentioning

confidence: 99%