+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-10454 | |||||||||
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Title | Campylobacter jejuni fliH deletion flagellar motor | |||||||||
Map data | Campylobacter jejuni %u0394fliH flagellar motor in situ subtomogram average. | |||||||||
Sample |
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Biological species | Campylobacter jejuni (Campylobacter) | |||||||||
Method | subtomogram averaging / cryo EM / Resolution: 43.0 Å | |||||||||
Authors | Matthews-Palmer T / Beeby M | |||||||||
Funding support | United Kingdom, 2 items
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Citation | Journal: mBio / Year: 2020 Title: Diversification of Campylobacter jejuni Flagellar C-Ring Composition Impacts Its Structure and Function in Motility, Flagellar Assembly, and Cellular Processes. Authors: Louie D Henderson / Teige R S Matthews-Palmer / Connor J Gulbronson / Deborah A Ribardo / Morgan Beeby / David R Hendrixson / Abstract: Bacterial flagella are reversible rotary motors that rotate external filaments for bacterial propulsion. Some flagellar motors have diversified by recruiting additional components that influence ...Bacterial flagella are reversible rotary motors that rotate external filaments for bacterial propulsion. Some flagellar motors have diversified by recruiting additional components that influence torque and rotation, but little is known about the possible diversification and evolution of core motor components. The mechanistic core of flagella is the cytoplasmic C ring, which functions as a rotor, directional switch, and assembly platform for the flagellar type III secretion system (fT3SS) ATPase. The C ring is composed of a ring of FliG proteins and a helical ring of surface presentation of antigen (SPOA) domains from the switch proteins FliM and one of two usually mutually exclusive paralogs, FliN or FliY. We investigated the composition, architecture, and function of the C ring of , which encodes FliG, FliM, and both FliY and FliN by a variety of interrogative approaches. We discovered a diversified C ring containing FliG, FliM, and both FliY, which functions as a classical FliN-like protein for flagellar assembly, and FliN, which has neofunctionalized into a structural role. Specific protein interactions drive the formation of a more complex heterooligomeric C-ring structure. We discovered that this complex C ring has additional cellular functions in polarly localizing FlhG for numerical regulation of flagellar biogenesis and spatial regulation of division. Furthermore, mutation of the C ring revealed a T3SS that was less dependent on its ATPase complex for assembly than were other systems. Our results highlight considerable evolved flagellar diversity that impacts motor output, biogenesis, and cellular processes in different species. The conserved core of bacterial flagellar motors reflects a shared evolutionary history that preserves the mechanisms essential for flagellar assembly, rotation, and directional switching. In this work, we describe an expanded and diversified set of core components in the flagellar C ring, the mechanistic core of the motor. Our work provides insight into how usually conserved core components may have diversified by gene duplication, enabling a division of labor of the ancestral protein between the two new proteins, acquisition of new roles in flagellar assembly and motility, and expansion of the function of the flagellum beyond motility, including spatial regulation of cell division and numerical control of flagellar biogenesis in Our results highlight that relatively small changes, such as gene duplications, can have substantial ramifications on the cellular roles of a molecular machine. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_10454.map.gz | 11.1 MB | EMDB map data format | |
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Header (meta data) | emd-10454-v30.xml emd-10454.xml | 10 KB 10 KB | Display Display | EMDB header |
Images | emd_10454.png | 83.3 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-10454 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-10454 | HTTPS FTP |
-Validation report
Summary document | emd_10454_validation.pdf.gz | 232.1 KB | Display | EMDB validaton report |
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Full document | emd_10454_full_validation.pdf.gz | 231.2 KB | Display | |
Data in XML | emd_10454_validation.xml.gz | 5.8 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10454 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10454 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_10454.map.gz / Format: CCP4 / Size: 12.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Campylobacter jejuni %u0394fliH flagellar motor in situ subtomogram average. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 8.28 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : Flagellar motor of Campylobacter jejuni in situ, fliH deletion strain.
Entire | Name: Flagellar motor of Campylobacter jejuni in situ, fliH deletion strain. |
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Components |
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-Supramolecule #1: Flagellar motor of Campylobacter jejuni in situ, fliH deletion strain.
Supramolecule | Name: Flagellar motor of Campylobacter jejuni in situ, fliH deletion strain. type: organelle_or_cellular_component / ID: 1 / Parent: 0 |
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Source (natural) | Organism: Campylobacter jejuni (Campylobacter) / Strain: 81-176 / Organelle: Flagellar motor / Location in cell: cell pole |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | subtomogram averaging |
Aggregation state | cell |
-Sample preparation
Buffer | pH: 7.3 |
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Grid | Model: UltrAuFoil / Material: GOLD / Mesh: 200 / Pretreatment - Type: GLOW DISCHARGE |
Vitrification | Cryogen name: ETHANE-PROPANE / Chamber humidity: 100 % / Chamber temperature: 20 K / Instrument: FEI VITROBOT MARK IV |
Details | Bacterial cell suspension mixed with 10nm gold fiducial nanoparticles. |
-Electron microscopy
Microscope | FEI TECNAI F20 |
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Image recording | Film or detector model: FEI FALCON II (4k x 4k) / Detector mode: INTEGRATING / Average electron dose: 120.0 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 100.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus min: 3.0 µm / Nominal magnification: 25000 |
Sample stage | Specimen holder model: GATAN 914 HIGH TILT LIQUID NITROGEN CRYO TRANSFER TOMOGRAPHY HOLDER Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Tecnai F20 / Image courtesy: FEI Company |
-Image processing
Final reconstruction | Applied symmetry - Point group: C17 (17 fold cyclic) / Resolution.type: BY AUTHOR / Resolution: 43.0 Å / Resolution method: FSC 0.5 CUT-OFF Details: Non-independent half-map FSC of unsymmetrised map used with 0.5 threshold cited as resolution. Number subtomograms used: 285 |
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Extraction | Number tomograms: 263 / Number images used: 285 / Method: manual selection / Software - Name: IMOD Details: reference-free alignment from manual initial co-ordinates & orientations. |
Final angle assignment | Type: NOT APPLICABLE / Software - Name: PEET |