1887

Abstract

Negatively stained influenza virions sometimes show irregular morphology and are often referred to as pleomorphic. However, this irregular morphology has not been visualized when ultrathin-section transmission and scanning electron microscopies are used. This study focused on the effects of ultracentrifugation on influenza A virion morphology, as negative staining often involves ultracentrifugation to concentrate or purify virions. The morphologies of unfixed, glutaraldehyde-fixed and osmium tetroxide-fixed virions were quantitatively compared before and after ultracentrifugation, and it was found that, without chemical fixation, approximately 30 % of virions were altered from oval to irregular shapes following ultracentrifugation. By contrast, most glutaraldehyde-fixed virions remained uniformly elliptical, even after ultracentrifugation. When a virus with an 11 aa deletion at the C terminus of its M2 cytoplasmic tail was ultracentrifuged, its morphology was appreciably deformed compared with that of the wild-type virus. These results demonstrate that the native morphology of influenza A virions is regular but is disrupted by ultracentrifugation, and that the cytoplasmic tail of M2 is important for virion integrity.

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2011-11-01
2021-10-23
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References

  1. Almeida J. D., Waterson A. P. 1967; Some observations on the envelope of an influenza virus. J Gen Microbiol 46:107–110[PubMed] [CrossRef]
    [Google Scholar]
  2. Almeida J. D., Waterson A. P., Drewe J. A. 1967; A morphological comparison of Bittner and influenza viruses. J Hyg (Lond) 65:467–474 [View Article][PubMed]
    [Google Scholar]
  3. Bächi T., Gerhard W., Lindenmann J., Mühlethaler K. 1969; Morphogenesis of influenza A virus in Ehrlich ascites tumor cells as revealed by thin-sectioning and freeze-etching. J Virol 4:769–776[PubMed]
    [Google Scholar]
  4. Booy F. P., Ruigrok R. W., van Bruggen E. F. 1985; Electron microscopy of influenza virus. A comparison of negatively stained and ice-embedded particles. J Mol Biol 184:667–676 [View Article][PubMed]
    [Google Scholar]
  5. Boulan E. R., Sabatini D. D. 1978; Asymmetric budding of viruses in epithelial monolayers: a model system for study of epithelial polarity. Proc Natl Acad Sci U S A 75:5071–5075 [View Article][PubMed]
    [Google Scholar]
  6. Bourmakina S. V., García-Sastre A. 2003; Reverse genetics studies on the filamentous morphology of influenza A virus. J Gen Virol 84:517–527 [View Article][PubMed]
    [Google Scholar]
  7. Bozzola J. J., Russell L. D. 1998; The mechanism of chemical fixation. In Electron Microscopy, 2nd edn. pp. 20–22 Edited by McKean B. L. Massachusetts: Jones and Bartlett Publishers;
    [Google Scholar]
  8. Burleigh L. M., Calder L. J., Skehel J. J., Steinhauer D. A. 2005; Influenza A viruses with mutations in the M1 helix six domain display a wide variety of morphological phenotypes. J Virol 79:1262–1270 [View Article][PubMed]
    [Google Scholar]
  9. Chen B. J., Leser G. P., Jackson D., Lamb R. A. 2008; The influenza virus M2 protein cytoplasmic tail interacts with the M1 protein and influences virus assembly at the site of virus budding. J Virol 82:10059–10070 [View Article][PubMed]
    [Google Scholar]
  10. Compans R. W., Dimmock N. J. 1969; An electron microscopic study of single-cycle infection of chick embryo fibroblasts by influenza virus. Virology 39:499–515 [View Article][PubMed]
    [Google Scholar]
  11. Elleman C. J., Barclay W. S. 2004; The M1 matrix protein controls the filamentous phenotype of influenza A virus. Virology 321:144–153 [View Article][PubMed]
    [Google Scholar]
  12. Fujiyoshi Y., Kume N. P., Sakata K., Sato S. B. 1994; Fine structure of influenza A virus observed by electron cryo-microscopy. EMBO J 13:318–326[PubMed]
    [Google Scholar]
  13. Harris A., Forouhar F., Qiu S., Sha B., Luo M. 2001; The crystal structure of the influenza matrix protein M1 at neutral pH: M1–M1 protein interfaces can rotate in the oligomeric structures of M1. Virology 289:34–44 [View Article][PubMed]
    [Google Scholar]
  14. Harris A., Cardone G., Winkler D. C., Heymann J. B., Brecher M., White J. M., Steven A. C. 2006; Influenza virus pleiomorphy characterized by cryoelectron tomography. Proc Natl Acad Sci U S A 103:19123–19127 [View Article][PubMed]
    [Google Scholar]
  15. Iwatsuki-Horimoto K., Horimoto T., Noda T., Kiso M., Maeda J., Watanabe S., Muramoto Y., Fujii K., Kawaoka Y. 2006; The cytoplasmic tail of the influenza A virus M2 protein plays a role in viral assembly. J Virol 80:5233–5240 [View Article][PubMed]
    [Google Scholar]
  16. Jin H., Leser G. P., Zhang J., Lamb R. A. 1997; Influenza virus hemagglutinin and neuraminidase cytoplasmic tails control particle shape. EMBO J 16:1236–1247 [View Article][PubMed]
    [Google Scholar]
  17. Le Ru A., Jacob D., Transfiguracion J., Ansorge S., Henry O., Kamen A. A. 2010; Scalable production of influenza virus in HEK-293 cells for efficient vaccine manufacturing. Vaccine 28:3661–3671 [View Article][PubMed]
    [Google Scholar]
  18. Morgan C., Rose H. M., Moore D. H. 1956; Structure and development of viruses observed in the electron microscope. III. Influenza virus. J Exp Med 104:171–182 [View Article][PubMed]
    [Google Scholar]
  19. Morgan C., Hsu K. C., Rifkind R. A., Knox A. W., Rose H. M. 1961; The application of ferritin-conjugated antibody to electron microscopic studies of influenza virus in infected cells. I. The cellular surface. J Exp Med 114:825–832 [View Article][PubMed]
    [Google Scholar]
  20. Nayak D. P., Hui E. K., Barman S. 2004; Assembly and budding of influenza virus. Virus Res 106:147–165 [View Article][PubMed]
    [Google Scholar]
  21. Nayak D. P., Balogun R. A., Yamada H., Zhou Z. H., Barman S. 2009; Influenza virus morphogenesis and budding. Virus Res 143:147–161 [View Article][PubMed]
    [Google Scholar]
  22. Nermut M. V., Frank H. 1971; Fine structure of influenza A2 (Singapore) as revealed by negative staining, freeze-drying and freeze-etching. J Gen Virol 10:37–51 [View Article][PubMed]
    [Google Scholar]
  23. Neumann G., Watanabe T., Ito H., Watanabe S., Goto H., Gao P., Hughes M., Perez D. R., Donis R. et al. 1999; Generation of influenza A viruses entirely from cloned cDNAs. Proc Natl Acad Sci U S A 96:9345–9350 [View Article][PubMed]
    [Google Scholar]
  24. Noda T., Sagara H., Yen A., Takada A., Kida H., Cheng R. H., Kawaoka Y. 2006; Architecture of ribonucleoprotein complexes in influenza A virus particles. Nature 439:490–492 [View Article][PubMed]
    [Google Scholar]
  25. Palese P., Shaw M. L. 2007; Orthomyxoviridae: the viruses and their replication. In Fields Virology, 5th edn. pp. 1647–1689 Edited by Knipe D. M., Howley P. M. Philadelphia: Lippincott Williams & Wilkins;
    [Google Scholar]
  26. Roberts P. C., Lamb R. A., Compans R. W. 1998; The M1 and M2 proteins of influenza A virus are important determinants in filamentous particle formation. Virology 240:127–137 [View Article][PubMed]
    [Google Scholar]
  27. Ruigrok R. W., Wrigley N. G., Calder L. J., Cusack S., Wharton S. A., Brown E. B., Skehel J. J. 1986; Electron microscopy of the low pH structure of influenza virus haemagglutinin. EMBO J 5:41–49[PubMed]
    [Google Scholar]
  28. Ruigrok R. W., Hewat E. A., Wade R. H. 1992; Low pH deforms the influenza virus envelope. J Gen Virol 73:995–998 [View Article][PubMed]
    [Google Scholar]
  29. Satoh H., Ueda T., Kobatake Y. 1985; Oscillations in cell shape and size during locomotion and in contractile activities of Physarum polycephalum, Dictyostelium discoideum, Amoeba proteus and macrophages. Exp Cell Res 156:79–90 [View Article][PubMed]
    [Google Scholar]
  30. Stevenson J. P., Biddle F. 1966; Pleomorphism of influenza virus particles under the electron microscope. Nature 212:619–621 [View Article][PubMed]
    [Google Scholar]
  31. Wrigley N. G. 1979; Electron microscopy of influenza virus. Br Med Bull 35:35–38[PubMed]
    [Google Scholar]
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