An atomic model of brome mosaic virus using direct electron detection and real-space optimization

Nat Commun. 2014 Sep 4:5:4808. doi: 10.1038/ncomms5808.

Abstract

Advances in electron cryo-microscopy have enabled structure determination of macromolecules at near-atomic resolution. However, structure determination, even using de novo methods, remains susceptible to model bias and overfitting. Here we describe a complete workflow for data acquisition, image processing, all-atom modelling and validation of brome mosaic virus, an RNA virus. Data were collected with a direct electron detector in integrating mode and an exposure beyond the traditional radiation damage limit. The final density map has a resolution of 3.8 Å as assessed by two independent data sets and maps. We used the map to derive an all-atom model with a newly implemented real-space optimization protocol. The validity of the model was verified by its match with the density map and a previous model from X-ray crystallography, as well as the internal consistency of models from independent maps. This study demonstrates a practical approach to obtain a rigorously validated atomic resolution electron cryo-microscopy structure.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bromovirus / ultrastructure*
  • Cryoelectron Microscopy / instrumentation
  • Cryoelectron Microscopy / methods*
  • Crystallography, X-Ray
  • Electrons*
  • Image Processing, Computer-Assisted / statistics & numerical data*
  • Models, Molecular
  • Protein Conformation

Associated data

  • PDB/3J7L
  • PDB/3J7M
  • PDB/3J7N