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Towards in cellulo virus crystallography

DOI: 10.1038/s41598-018-21693-3 DOI Help

Authors: Helen M. E. Duyvesteyn (University of Oxford; Diamond Light Source) , Helen M. Ginn (University of Oxford; Diamond Light Source) , Maija K. Pietila (University of Helsinki) , Armin Wagner (Diamond Light Source) , Johan Hattne (Lawrence Berkeley National Laboratory) , Jonathan M. Grimes (University of Oxford) , Elina Hirvonen (University of Helsinki) , Gwyndaf Evans (Diamond Light Source) , Marie-laure Parsy (University of Oxford) , Nicholas K. Sauter (Lawrence Berkeley National Laboratory) , Aaron S. Brewster (Lawrence Berkeley National Laboratory) , Juha Huiskonen (University of Oxford; University of Helsinki) , David I. Stuart (University of Oxford; Diamond Light Source) , Geoff Sutton (University of Oxford) , Dennis H. Bamford (University of Helsinki)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Scientific Reports , VOL 8

State: Published (Approved)
Published: February 2018

Open Access Open Access

Abstract: 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.

Journal Keywords: Bacteriophages; Nanocrystallography

Subject Areas: Biology and Bio-materials, Medicine

Facility: SLAC National Accelerator Laboratory