Journal: Nat Microbiol / Year: 2019 Title: Structural dynamics of bacteriophage P22 infection initiation revealed by cryo-electron tomography. Authors: Chunyan Wang / Jiagang Tu / Jun Liu / Ian J Molineux / Abstract: For successful infection, bacteriophages must overcome multiple barriers to transport their genome and proteins across the bacterial cell envelope. We use cryo-electron tomography to study the ...For successful infection, bacteriophages must overcome multiple barriers to transport their genome and proteins across the bacterial cell envelope. We use cryo-electron tomography to study the infection initiation of phage P22 in Salmonella enterica serovar Typhimurium, revealing how a channel forms to allow genome translocation into the cytoplasm. Our results show free phages that initially attach obliquely to the cell through interactions between the O antigen and two of the six tailspikes; the tail needle also abuts the cell surface. The virion then orients perpendicularly and the needle penetrates the outer membrane. The needle is released and the internal head protein gp7* is ejected and assembles into an extracellular channel that extends from the gp10 baseplate to the cell surface. A second protein, gp20, is ejected and assembles into a structure that extends the extracellular channel across the outer membrane into the periplasm. Insertion of the third ejected protein, gp16, into the cytoplasmic membrane probably completes the overall trans-envelope channel into the cytoplasm. Construction of a trans-envelope channel is an essential step during infection of Gram-negative bacteria by all short-tailed phages, because such virions cannot directly deliver their genome into the cell cytoplasm.
History
Deposition
Jul 30, 2018
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Header (metadata) release
Aug 22, 2018
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Map release
Apr 3, 2019
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Update
Jun 5, 2019
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Current status
Jun 5, 2019
Processing site: RCSB / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
Download / File: emd_9009.map.gz / Format: CCP4 / Size: 178 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotation
Channel structure formed by Gp7 defective mutant of bacteriophage P22 in Salmonella
Voxel size
X=Y=Z: 4.5 Å
Density
Contour Level
By AUTHOR: 0.08 / Movie #1: 0.08
Minimum - Maximum
-0.58579206 - 0.5763105
Average (Standard dev.)
-0.000000000003131 (±0.038722493)
Symmetry
Space group: 1
Details
EMDB XML:
Map geometry
Axis order
X
Y
Z
Origin
-180
-180
-180
Dimensions
360
360
360
Spacing
360
360
360
Cell
A=B=C: 1620.0 Å α=β=γ: 90.0 °
CCP4 map header:
mode
Image stored as Reals
Å/pix. X/Y/Z
4.5
4.5
4.5
M x/y/z
360
360
360
origin x/y/z
0.000
0.000
0.000
length x/y/z
1620.000
1620.000
1620.000
α/β/γ
90.000
90.000
90.000
MAP C/R/S
1
2
3
start NC/NR/NS
-180
-180
-180
NC/NR/NS
360
360
360
D min/max/mean
-0.586
0.576
-0.000
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Supplemental data
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Sample components
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Entire : Channel structure formed by Gp7-defective mutant of bacteriophage...
Entire
Name: Channel structure formed by Gp7-defective mutant of bacteriophage P22 in Salmonella using cryoelectron tomography
Components
Complex: Channel structure formed by Gp7-defective mutant of bacteriophage P22 in Salmonella using cryoelectron tomography
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Supramolecule #1: Channel structure formed by Gp7-defective mutant of bacteriophage...
Supramolecule
Name: Channel structure formed by Gp7-defective mutant of bacteriophage P22 in Salmonella using cryoelectron tomography type: complex / ID: 1 / Parent: 0
Source (natural)
Organism: Enterobacteria phage P22 (virus)
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Experimental details
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Structure determination
Method
cryo EM
Processing
subtomogram averaging
Aggregation state
cell
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Sample preparation
Buffer
pH: 5
Grid
Details: unspecified
Vitrification
Cryogen name: ETHANE
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Electron microscopy
Microscope
FEI POLARA 300
Image recording
Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 1.5 e/Å2
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron optics
Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD
Experimental equipment
Model: Tecnai Polara / Image courtesy: FEI Company
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Image processing
Final reconstruction
Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 28.0 Å / Resolution method: FSC 0.143 CUT-OFF / Number subtomograms used: 4319
Extraction
Number tomograms: 149 / Number images used: 6636
Final angle assignment
Type: ANGULAR RECONSTITUTION
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