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2018 ; 8
(1
): 3771
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Towards in cellulo virus crystallography
#MMPMID29491457
Duyvesteyn HME
; Ginn HM
; Pietilä MK
; Wagner A
; Hattne J
; Grimes JM
; Hirvonen E
; Evans G
; Parsy ML
; Sauter NK
; Brewster AS
; Huiskonen JT
; Stuart DI
; Sutton G
; Bamford DH
Sci Rep
2018[Feb]; 8
(1
): 3771
PMID29491457
show ga
Viruses are a significant threat to both human health and the economy, and there
is an urgent need for novel anti-viral drugs and vaccines. High-resolution viral
structures inform our understanding of the virosphere, and inspire novel
therapies. Here we present a method of obtaining such structural information that
avoids potentially disruptive handling, by collecting diffraction data from
intact infected cells. We identify a suitable combination of cell type and virus
to accumulate particles in the cells, establish a suitable time point where most
cells contain virus condensates and use electron microscopy to demonstrate that
these are ordered crystalline arrays of empty capsids. We then use an X-ray free
electron laser to provide extremely bright illumination of sub-micron
intracellular condensates of bacteriophage phiX174 inside living Escherichia coli
at room temperature. We have been able to collect low resolution diffraction
data. Despite the limited resolution and completeness of these initial data, due
to a far from optimal experimental setup, we have used novel methodology to
determine a putative space group, unit cell dimensions, particle packing and
likely maturation state of the particles.
|*Crystallography, X-Ray
[MESH]
|Bacteriophage phi X 174/*chemistry/physiology
[MESH]
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