Journal: Nature / Year: 2013 Title: Visualizing virus assembly intermediates inside marine cyanobacteria. Authors: Wei Dai / Caroline Fu / Desislava Raytcheva / John Flanagan / Htet A Khant / Xiangan Liu / Ryan H Rochat / Cameron Haase-Pettingell / Jacqueline Piret / Steve J Ludtke / Kuniaki Nagayama / ...Authors: Wei Dai / Caroline Fu / Desislava Raytcheva / John Flanagan / Htet A Khant / Xiangan Liu / Ryan H Rochat / Cameron Haase-Pettingell / Jacqueline Piret / Steve J Ludtke / Kuniaki Nagayama / Michael F Schmid / Jonathan A King / Wah Chiu / Abstract: Cyanobacteria are photosynthetic organisms responsible for ∼25% of organic carbon fixation on the Earth. These bacteria began to convert solar energy and carbon dioxide into bioenergy and oxygen ...Cyanobacteria are photosynthetic organisms responsible for ∼25% of organic carbon fixation on the Earth. These bacteria began to convert solar energy and carbon dioxide into bioenergy and oxygen more than two billion years ago. Cyanophages, which infect these bacteria, have an important role in regulating the marine ecosystem by controlling cyanobacteria community organization and mediating lateral gene transfer. Here we visualize the maturation process of cyanophage Syn5 inside its host cell, Synechococcus, using Zernike phase contrast electron cryo-tomography (cryoET). This imaging modality yields dramatic enhancement of image contrast over conventional cryoET and thus facilitates the direct identification of subcellular components, including thylakoid membranes, carboxysomes and polyribosomes, as well as phages, inside the congested cytosol of the infected cell. By correlating the structural features and relative abundance of viral progeny within cells at different stages of infection, we identify distinct Syn5 assembly intermediates. Our results indicate that the procapsid releases scaffolding proteins and expands its volume at an early stage of genome packaging. Later in the assembly process, we detected full particles with a tail either with or without an additional horn. The morphogenetic pathway we describe here is highly conserved and was probably established long before that of double-stranded DNA viruses infecting more complex organisms.
History
Deposition
Aug 13, 2013
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Header (metadata) release
Aug 28, 2013
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Map release
Oct 9, 2013
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Update
Nov 6, 2013
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Current status
Nov 6, 2013
Processing site: RCSB / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 85 K / Instrument: FEI VITROBOT MARK III / Method: Blot for 3 seconds before plunging
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Electron microscopy
Microscope
JEOL 2200FS
Temperature
Min: 93 K / Max: 105 K / Average: 103 K
Alignment procedure
Legacy - Astigmatism: Objective lens astigmatism was corrected at 100,000 times magnification
Specialist optics
Energy filter - Name: in-column GIF energy filter / Energy filter - Lower energy threshold: 15.0 eV / Energy filter - Upper energy threshold: 20.0 eV
Date
Jun 1, 2011
Image recording
Category: CCD / Film or detector model: GATAN ULTRASCAN 4000 (4k x 4k) / Number real images: 42 / Average electron dose: 1 e/Å2 / Camera length: 1200 / Bits/pixel: 16
Electron beam
Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
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