National Institutes of Health/National Institute of General Medical Sciences
U24GM116789-03
United States
AAAS Marion Mason Milligan Award for Women in the Chemical Sciences
United States
National Institutes of Health/National Human Genome Research Institute
GM110185
United States
NVIDIA GPU grant program
United States
Burroughs Wellcome Fund Investigators in the Pathogenicity of Disease
United States
Citation
Journal: Cell / Year: 2020 Title: Structural and Proteomic Characterization of the Initiation of Giant Virus Infection. Authors: Jason R Schrad / Jônatas S Abrahão / Juliana R Cortines / Kristin N Parent / Abstract: Since their discovery, giant viruses have expanded our understanding of the principles of virology. Due to their gargantuan size and complexity, little is known about the life cycles of these viruses. ...Since their discovery, giant viruses have expanded our understanding of the principles of virology. Due to their gargantuan size and complexity, little is known about the life cycles of these viruses. To answer outstanding questions regarding giant virus infection mechanisms, we set out to determine biomolecular conditions that promote giant virus genome release. We generated four infection intermediates in Samba virus (Mimivirus genus, lineage A) as visualized by cryoelectron microscopy (cryo-EM), cryoelectron tomography (cryo-ET), and scanning electron microscopy (SEM). Each of these four intermediates reflects similar morphology to a stage that occurs in vivo. We show that these genome release stages are conserved in other mimiviruses. Finally, we identified proteins that are released from Samba and newly discovered Tupanvirus through differential mass spectrometry. Our work revealed the molecular forces that trigger infection are conserved among disparate giant viruses. This study is also the first to identify specific proteins released during the initial stages of giant virus infection.
Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Average exposure time: 5.0 sec. / Average electron dose: 2.5 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
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Image processing
Final reconstruction
Algorithm: SIMULTANEOUS ITERATIVE (SIRT) / Software - Name: IMOD (ver. 4.7.5) / Number images used: 51
Details
MotionCor processing to create final aligned images
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