Structural studies of viruses by electron cryomicroscopy

  • M. F. Schmid
  • B. V. V. Prasad
  • W. Chiu
Conference paper
Part of the Archives of Virology Supplementum book series (ARCHIVES SUPPL, volume 9)


Electron cryomicroscopy is a unique biophysical technique for studying molecular structures of viruses which are difficult to analyze by x-ray diffraction. The structural information derived from the low resolution reconstructions of viruses has so far been useful to understand various functional properties of the viruses such as antibody neutralization, receptor binding and assembly. Electron cryomicroscopy has enabled the visualization of the four core alpha helices of the coat protein in tobacco mosaic virus. This represents the highest resolution detail of a virus studied by electron cryomicroscopy. The prospects of attaining similar resolution beyond 10 Å for spherical viruses as well are encouraging, with newly available instrumentation, data collection and processing procedures.


Rota Virus Tobacco Mosaic Virus Tomato Bushy Stunt Virus Image Processing Step Icosahedral Virus 
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.


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  1. 1.
    Adrian M, Dubochet J, Lepault J, McDowall AW (1984) Cryo-electron microscopy of viruses. Nature 308: 32–36PubMedCrossRefGoogle Scholar
  2. 2.
    Aldroubi A, Trus BL, Unser M, Booy FP, Steven AC (1992) Magnification mismatches between micrographs: corrective procedures and implications for structural analysis. Ultramicroscopy 46: 175–188PubMedCrossRefGoogle Scholar
  3. 3.
    Amos LA, Klug A (1975) Three-dimensional image reconstructions of the contractile tail of T4 bacteriophage. J Mol Biol 99: 51–73PubMedCrossRefGoogle Scholar
  4. 4.
    Baker TS, Cheng RH, Johnson JE, Olson NH, Wang GJ, Schmidt TJ (1992) Organized packing of RNA inside viruses as revealed by cryo-electron microscopy and x-ray diffraction analysis. In: 50th Ann Electr Micros Soc Amer. Boston, San Francisco Press, pp 454–455Google Scholar
  5. 5.
    Bloomer AC, Champness JN, Bricogne G, Staden R, Klug A (1978) Protein disk of tobacco mosaic virus at 2.8 Å resolution showing the interactions within and between subunits. Nature 276: 362–368PubMedCrossRefGoogle Scholar
  6. 6.
    Booy F, Trus BL, Newcomb WW, Brown JC, Serwer P, Steven AC (1992) Organization of dsDNA in icosahedral virus capsids. In: 50th Ann Electr Microsc Sco Amer. Boston, San Francisco Press, pp 452–453Google Scholar
  7. 7.
    Caspar DLD, Namba K (1990) Switching in the self-assembly of tobacco mosaic virus. Adv Biophys 26: 157–185PubMedCrossRefGoogle Scholar
  8. 8.
    Cheng RH (1992) Correlation of cryo-electron microscopic and x-ray data and compensation of the contrast transfer function. In: Proc 50th Ann Meeting of Electr Microsc Soc Amer. Boston, San Francisco Press, pp 996–997Google Scholar
  9. 9.
    Chiu W (1986) Electron microscopy of frozen, hydrated biological specimens. Annu Rev Biophys Biomol Struct 15: 237–257CrossRefGoogle Scholar
  10. 10.
    Chiu W (1993) What does electron cryomicroscopy provide that X-ray crystallography and NMR spectroscopy cannot? Annu Rev Biophys Biomol Struct 22: 233–255PubMedCrossRefGoogle Scholar
  11. 11.
    Chiu W, Schmid MF, Prasad BVV (1993) Teaching electron diffraction and imaging of macromolecules. Biophys J 64: 397–402Google Scholar
  12. 12.
    Crowther RA (1971) Procedures for three-dimensional reconstruction of spherical viruses by Fourier synthesis from electron micrographs. Phil Trans Roy Soc London Ser B 261: 221–230CrossRefGoogle Scholar
  13. 13.
    Crowther RA, DeRosier DJ, Klug A (1970) The reconstruction of a three- dimensional structure from projections and its application to electron microscopy. Proc Roy Soc London 317: 319–340CrossRefGoogle Scholar
  14. 14.
    DeRosier DJ, Klug A (1968) Reconstruction of three-dimensional structures from electrons micrographs. Nature 217: 130–134CrossRefGoogle Scholar
  15. 15.
    DeRosier DJ, Moore PB (1970) Reconstruction of three-dimensional images from electron micrographs of structures with helical symmetry. J Mol Biol 52: 355–369CrossRefGoogle Scholar
  16. 16.
    Downing KH, Chiu W (1990) Cold stage design for high resolution electron microscopy of biological materials. J Electron Microsc (Tokyo) 3: 213–226Google Scholar
  17. 17.
    Dubochet J, Adrian M, Chang JJ, Homo JC, Lepault J, McDowall AW, Schultz P (1988) Cryo-electron microscopy of vitrified specimens. Q Rev Biophys 21: 129–228PubMedCrossRefGoogle Scholar
  18. 18.
    Frank J, Radermacher M (1992) Three-dimensional reconstruction of single particles negatively stained or in vitreous ice. Ultramicroscopy 46: 241–262PubMedCrossRefGoogle Scholar
  19. 19.
    Fuller SD (1987) The T = 4 envelope of Sindbis virus is organized by interactions with a complementary T = 3 capsid. Cell 48: 923–934PubMedCrossRefGoogle Scholar
  20. 20.
    Harrison SC, Olson A, Schutt CE, Winkler FK, Bricogne G (1978) Tomato bushy stunt virus at 2.9 Å resolution. Nature 276: 368–373PubMedCrossRefGoogle Scholar
  21. 21.
    Hogle JM, Maeda A, Harrison SC (1986) The structure and assembly of turnip crinkle virus I: X-ray crystallographic analysis at 3.2 Å. J Mol Biol 191: 625–638PubMedCrossRefGoogle Scholar
  22. 22.
    Jeng TW, Crowther RA, Stubbs G, Chiu W (1989) Visualization of alpha-helices in tobacco mosaic virus by cryo-electron microscopy. J Mol Biol 205: 251–257PubMedCrossRefGoogle Scholar
  23. 23.
    Jeng TW, Talmon Y, Chiu W (1988) Containment system for the preparation of vitrified-hydrated virus specimens. J Electron Microsc Tech 8: 343–348PubMedCrossRefGoogle Scholar
  24. 24.
    Klug A (1983) From macromolecules to biological assemblies. Bioscience Reports 3: 395–430PubMedCrossRefGoogle Scholar
  25. 25.
    Levine AJ (1992) Viruses Scientific American Library. W.H. Freeman, New YorkGoogle Scholar
  26. 26.
    Namba K, Stubbs G (1986) Structure of TMV at 3.6 Å resolution: implications for assembly. Science 231: 1401–1406PubMedCrossRefGoogle Scholar
  27. 27.
    Olson NH, Kolatkar PR, Oliveira MA, Cheng RH, Greve JM, McClelland A, Baker TS, Rossmann MG (1993) Structure of a human rhinovirus complexed with its receptor molecule. Proc Natl Acad Sci USA 90: 507–511PubMedCrossRefGoogle Scholar
  28. 28.
    Prasad BVV, Burns JW, Marietta E, Estes MK, Chiu W (1990) Localization of VP4 neutralization sites in rotavirus by three-dimensional cryo-electron microscopy. Nature 343: 476–479PubMedCrossRefGoogle Scholar
  29. 29.
    Prasad BVV, Prevelige P, Marietta E, Chen R, Thomas D, King J, Chiu W (1993) Three-dimensional transformation of capsids associated with genome packaging in a bacterial virus. J Mol Biol 231: 65–74PubMedCrossRefGoogle Scholar
  30. 30.
    Prasad BVV, Matson DO, Smith AW (1993) Three-dimensional structure of a calicivirus (submitted)Google Scholar
  31. 31.
    Stewart PL, Burnett RM, Cyrklaff M, Fuller SD (1991) Image reconstruction reveals the complex molecular organization of adenovirus. Cell 67: 145–154PubMedCrossRefGoogle Scholar
  32. 32.
    Wang G, Porta C, Chen Z, Baker T, Johnson JE (1992) Identification of a Fab interaction footprint site on an icosahedral virus by cryoelectron microscopy and x-ray crystallography. Nature 355: 275–278PubMedCrossRefGoogle Scholar
  33. 33.
    Zhou ZH, Chiu W (1993) Prospects for using an IVEM with a FEG for imaging macromolecules towards atomic resolution. Ultramicroscopy 49: 407–416PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • M. F. Schmid
    • 1
  • B. V. V. Prasad
  • W. Chiu
  1. 1.Verna and Marrs McLean Department of Biochemistry, The W. M. Keck Center for Computational BiologyBaylor College of MedicineHoustonUSA

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