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Reconstructing Virus Structures from Nanometer to Near-Atomic Resolutions with Cryo-Electron Microscopy and Tomography

  • Juan Chang
  • Xiangan Liu
  • Ryan H. Rochat
  • Matthew L. Baker
  • Wah ChiuEmail author
Chapter
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 726)

Abstract

The past few decades have seen tremendous advances in single-particle electron ­cryo-microscopy (cryo-EM). The field has matured to the point that near-atomic resolution density maps can be generated for icosahedral viruses without the need for crystallization. In parallel, substantial progress has been made in determining the structures of nonicosahedrally arranged proteins in viruses by employing either single-particle cryo-EM or cryo-electron tomography (cryo-ET). Implicit in this course have been the availability of a new generation of electron cryo-microscopes and the development of the computational tools that are essential for generating these maps and models. This methodology has enabled structural biologists to analyze structures in increasing detail for virus particles that are in different morphogenetic states. Furthermore, electron imaging of frozen, hydrated cells, in the process of being infected by viruses, has also opened up a new avenue for studying virus structures “in situ”. Here we present the common techniques used to acquire and process cryo-EM and cryo-ET data and discuss their implications for structural virology both now and in the future.

Keywords

Particle Image Fourier Space Icosahedral Symmetry Icosahedral Virus Contrast Transfer Function 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Abbreviations

1-D

One dimension; one dimensional

2-D

Two dimensions; two dimensional

3-D

Three dimensions; three dimensional

CCD

Charge-coupled device

Cryo-EM

Electron cryo-microscopy

Cryo-ET

Electron cryo-tomography

CT

Computed tomography

CTF

Contrast transfer function

EM

Electron microscope; electron microscopy

EMDB

Electron Microscopy Data Bank (http://www.emdatabank.org)

FSC

Fourier shell correlation

FT

Fourier transform

GUI

Graphical user interface

SNR

Signal-to-noise ratio

SSE

Secondary structural elements

WPOA

Weak phase object approximation

ZPC

Zernike phase contrast

Notes

Acknowledgments

JC, XL, RHR and MLB contributed equally to this work. This work has been supported by grants from NIH (P41RR002250, R01AI0175208, and R01GM079429) and Robert Welch Foundation (Q1242). RHR is supported by the NIH training grants (GM07330 through the MSTP, T15LM007093 through the Gulf Coast Consortia). We thank Dr. Frazer Rixon for assistance with manuscript preparation.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Juan Chang
    • 1
  • Xiangan Liu
    • 1
  • Ryan H. Rochat
    • 1
    • 2
  • Matthew L. Baker
    • 1
  • Wah Chiu
    • 1
    • 2
    Email author
  1. 1.National Center for Macromolecular Imaging, Verna and Marrs McLean Department of Biochemistry and Molecular BiologyBaylor College of MedicineHoustonUSA
  2. 2.Graduate Program in Structural and Computational Biology and Molecular BiophysicsBaylor College of MedicineHoustonUSA

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