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Viruses at High Resolution

VP6 at 3.8 Å

We are collaborating with Stephen Harrison (HHMI, Harvard Medical School) to use virus particles as test specimens to develop better single particle image-processing methods. Virus particles have a high degree of symmetry and are stable in an aqueous solution, making them ideal for EM imaging.

We have used rotavirus particles to determine a structure of several of the capsid proteins to a resolution of 3.8 Å (Figure) [1][2]. Rotavirus is also a good particle to investigate current limitations in cryo-EM, such as beam-induced motion, and to develop and test new image processing methods, such as motion correction [3][4][5][6][7][8].

Furthermore, we use helical viruses, such as TMV [9], to test new algorithms for helical particle processing.


References

  1. Zhang, X, Settembre E, Xu C, Dormitzer PR, Bellamy R, Harrison SC, Grigorieff N.  2008.  Near-atomic resolution using electron cryomicroscopy and single-particle reconstruction. Proc Natl Acad Sci U S A. 105:1867-72.
  2. Chen, JZ, Settembre EC, Aoki ST, Zhang X, Bellamy AR, Dormitzer PR, Harrison SC, Grigorieff N.  2009.  Molecular interactions in rotavirus assembly and uncoating seen by high-resolution cryo-EM. Proc Natl Acad Sci U S A. 106:10644-48.
  3. Henderson, R, Chen S, Chen JZ, Grigorieff N, Passmore LA, Ciccarelli L, Rubinstein JL, Crowther RA, Stewart PL, Rosenthal PB.  2011.  Tilt-pair analysis of images from a range of different specimens in single-particle electron cryomicroscopy. J Mol Biol. 413:1028-46.
  4. Brilot, AF, Chen JZ, Cheng A, Pan J, Harrison SC, Potter CS, Carragher B, Henderson R, Grigorieff N.  2012.  Beam-induced motion of vitrified specimen on holey carbon film.. J Struct Biol. 177:630-637.
  5. Campbell, MG, Cheng A, Brilot AF, Moeller A, Lyumkis D, Veesler D, Pan J, Harrison SC, Potter CS, Carragher B et al..  2012.  Movies of ice-embedded particles enhance resolution in electron cryo-microscopy. Structure. 20:1823–1828.
  6. Grant, T, Grigorieff N.  2015.  Measuring the optimal exposure for single particle cryo-EM using a 2.6 Å reconstruction of rotavirus VP6. eLife. 4(e06980):1-19.
  7. Grant, T, Grigorieff N.  2015.  Automatic estimation and correction of anisotropic magnification distortion in electron microscopes. J Struct Biol. 192:204–208.
  8. Rickgauer, JP, Grigorieff N, Denk W.  2017.  Single-protein detection in crowded molecular environments in cryo-EM images. eLife. 6(e25648):1-22.
  9. Sachse, C, Chen JZ, Coureux PD, Stroupe ME, Fändrich M, Grigorieff N.  2007.  High-resolution electron microscopy of helical specimens: a fresh look at tobacco mosaic virus. J Mol Biol. 371:812-35.