|Entry||Database: EMDB / ID: 5217|
|Title||Visualizing the structural changes of bacteriophage epsilon15 and its Salmonella host during infection|
|Map data||Empty capsid attached to cell|
|Sample||Bacteriophage epsilon15, Salmonella anatum:|
|Keywords||virus / infection / Salmonella / tomography / bacteriophage|
|Method||subtomogram averaging / cryo EM|
|Authors||Chang JT / Schmid MF / Haase-Pettingell C / Weigele PR / King JA / Chiu W|
|Citation||Journal: J. Mol. Biol. / Year: 2010|
Title: Visualizing the structural changes of bacteriophage Epsilon15 and its Salmonella host during infection.
Authors: Juan T Chang / Michael F Schmid / Cameron Haase-Pettingell / Peter R Weigele / Jonathan A King / Wah Chiu
Abstract: The efficient mechanism by which double-stranded DNA bacteriophages deliver their chromosome across the outer membrane, cell wall, and inner membrane of Gram-negative bacteria remains obscure. ...The efficient mechanism by which double-stranded DNA bacteriophages deliver their chromosome across the outer membrane, cell wall, and inner membrane of Gram-negative bacteria remains obscure. Advances in single-particle electron cryomicroscopy have recently revealed details of the organization of the DNA injection apparatus within the mature virion for various bacteriophages, including epsilon15 (ɛ15) and P-SSP7. We have used electron cryotomography and three-dimensional subvolume averaging to capture snapshots of ɛ15 infecting its host Salmonella anatum. These structures suggest the following stages of infection. In the first stage, the tailspikes of ɛ15 attach to the surface of the host cell. Next, ɛ15's tail hub attaches to a putative cell receptor and establishes a tunnel through which the injection core proteins behind the portal exit the virion. A tube spanning the periplasmic space is formed for viral DNA passage, presumably from the rearrangement of core proteins or from cellular components. This tube would direct the DNA into the cytoplasm and protect it from periplasmic nucleases. Once the DNA has been injected into the cell, the tube and portal seals, and the empty bacteriophage remains at the cell surface.
|Date||Deposition: Jul 22, 2010 / Header (metadata) release: Aug 4, 2010 / Map release: Oct 11, 2010 / Last update: Sep 12, 2011|
|Structure viewer||EM map: |
Downloads & links
|File||emd_5217.map.gz (map file in CCP4 format, 27649 KB)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 10.8 Å|
CCP4 map header:
-Entire Bacteriophage epsilon15, Salmonella anatum
|Entire||Name: Bacteriophage epsilon15, Salmonella anatum / Number of components: 2|
-Component #1: virus, epsilon15
|Virus||Name: epsilon15Epsilon 15 / a.k.a: epsilon15Epsilon 15 / Class: VIRION / Empty: No / Enveloped: No / Isolate: STRAIN|
|Species||Species: epsilon15 (bacteriophage)|
|Source (natural)||Host Species: Salmonella enterica subsp. enterica serovar Anatum|
Host category: BACTERIA(EUBACTERIA)
|Specimen||Specimen state: particle / Method: cryo EM|
|Support film||200 mesh copper grid|
|Vitrification||Instrument: FEI VITROBOT / Cryogen name: ETHANE / Temperature: 93 K / Humidity: 100 % / Details: Vitrification instrument: Vitrobot|
-Electron microscopy imaging
|Imaging||Microscope: JEOL 3200FSC / Date: Sep 16, 2006|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 65 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Magnification: 20000 X (nominal) / Imaging mode: BRIGHT FIELD / Defocus: 6000 - 9000 nm / Energy filter: JEM3200FSC / Energy window: 0-20 eV|
|Specimen Holder||Holder: Eucentric / Model: JEOL 3200FSC CRYOHOLDER / Temperature: 93 K ( 93 - 93 K)|
|Camera||Detector: GENERIC CCD|
|Processing||Method: subtomogram averaging|
|3D reconstruction||Details: The 3D reconstructions were performed with IMOD using gold as fiducial markers. Subvolumes, each containing a single particle, were computationally extracted from the reconstructions, aligned to a reference model, and averaged together.|
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