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Towards in cellulo virus crystallography

  • Author(s): 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
  • et al.

Published Web Location

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5830620/
No data is associated with this publication.
Abstract

© 2018 The Author(s). 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.

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