|Entry||Database: EMDB / ID: EMD-0634|
|Title||Helicobacter pylori Cag T4SS innermembrane complex|
|Biological species||Helicobacter pylori (bacteria)|
|Method||subtomogram averaging / cryo EM / Resolution: 47.8 Å|
|Authors||Hu B / Christie PJ|
|Citation||Journal: MBio / Year: 2019|
Title: Molecular Architecture of the Helicobacter pylori Cag Type IV Secretion System.
Authors: Bo Hu / Pratick Khara / Liqiang Song / Aung Soe Lin / Arwen E Frick-Cheng / M Lorena Harvey / Timothy L Cover / Peter J Christie /
Abstract: colonizes about half of humans worldwide, and its presence in the gastric mucosa is associated with an increased risk of gastric adenocarcinoma, gastric lymphoma, and peptic ulcer disease. strains ... colonizes about half of humans worldwide, and its presence in the gastric mucosa is associated with an increased risk of gastric adenocarcinoma, gastric lymphoma, and peptic ulcer disease. strains carrying the pathogenicity island (PAI) are associated with increased risk of disease progression. The PAI encodes the Cag type IV secretion system (Cag), which delivers the CagA oncoprotein and other effector molecules into human gastric epithelial cells. We visualized structures of native and mutant Cag machines on the cell envelope by cryoelectron tomography. Individual cells contain multiple Cag nanomachines, each composed of a wheel-shaped outer membrane complex (OMC) with 14-fold symmetry and an inner membrane complex (IMC) with 6-fold symmetry. CagX, CagY, and CagM are required for assembly of the OMC, whereas strains lacking Cag3 and CagT produce outer membrane complexes lacking peripheral components. The IMC, which has never been visualized in detail, is configured as six tiers in cross-section view and three concentric rings surrounding a central channel in end-on view. The IMC contains three T4SS ATPases: (i) VirB4-like CagE, arranged as a hexamer of dimers at the channel entrance; (ii) a hexamer of VirB11-like Cagα, docked at the base of the CagE hexamer; and (iii) VirD4-like Cagβ and other unspecified Cag subunits, associated with the stacked CagE/Cagα complex and forming the outermost rings. The Cag and recently solved Dot/Icm system comprise new structural prototypes for the T4SS superfamily. Bacterial type IV secretion systems (T4SSs) have been phylogenetically grouped into two subfamilies. The T4ASSs, represented by the VirB/VirD4, include "minimized" machines assembled from 12 VirB- and VirD4-like subunits and compositionally larger systems such as the Cag T4BSSs encompass systems closely related in subunit composition to the Dot/Icm Here, we present structures of native and mutant Cag machines determined by cryoelectron tomography. We identify distinct outer and inner membrane complexes and, for the first time, visualize structural contributions of all three "signature" ATPases of T4SSs at the cytoplasmic entrance of the translocation channel. Despite their evolutionary divergence, the Cag aligns structurally much more closely to the Dot/Icm than an available VirB/VirD4 subcomplex. Our findings highlight the diversity of T4SSs and suggest a structural classification scheme in which T4SSs are grouped as minimized VirB/VirD4-like or larger Cag-like and Dot/Icm-like systems.
|Date||Deposition: Mar 6, 2019 / Header (metadata) release: May 29, 2019 / Map release: May 29, 2019 / Update: May 29, 2019|
|Structure viewer||EM map: |
Downloads & links
|File||Download / File: emd_0634.map.gz / Format: CCP4 / Size: 2 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 9 Å|
|Symmetry||Space group: 1|
CCP4 map header:
-Entire Cag T4SS
|Entire||Name: Cag T4SS / Number of components: 1|
-Component #1: cellular-component, Cag T4SS
|Cellular-component||Name: Cag T4SS / Recombinant expression: No|
|Source||Species: Helicobacter pylori (bacteria)|
|Specimen||Specimen state: Cell / Method: cryo EM|
|Sample solution||pH: 7|
|Vitrification||Cryogen name: ETHANE|
-Electron microscopy imaging
Model: Tecnai Polara / Image courtesy: FEI Company
|Imaging||Microscope: FEI POLARA 300|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 1.8 e/Å2 / Illumination mode: OTHER|
|Lens||Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: OTHER|
|Camera||Detector: GATAN K2 SUMMIT (4k x 4k)|
|Processing||Method: subtomogram averaging / Applied symmetry: C6 (6 fold cyclic) / Number of subtomograms: 1280|
|3D reconstruction||Resolution: 47.8 Å / Resolution method: FSC 0.5 CUT-OFF|
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