1887

Abstract

SUMMARY

We have determined the nucleotide sequences of the capsid, E3, E2 and 6K genes of the avirulent Semliki Forest virus variant A774 (SFV A7). The sequence analysis revealed a nucleotide identity of 98% for capsid, 98% for E3, 97% for E2 and 98% for 6K genes, as compared with the prototype SFV strain L10. At the protein level, the capsid and E3 polypeptides of SFV A7 both exhibited two amino acid substitutions, whereas point mutations in the 6K gene did not alter the amino acid sequence. In the E2 gene of SFV A7, seven of the 34 point mutations led to an amino acid difference as compared with the L10 strain. Replacement of the E2 glycoprotein gene of the virulent SFV4 clone with the corresponding region of SFV A7 resulted in a new plasmid construct, pME2, that gave rise to infectious virus CME2. CME2 and SFV4 replicated similarly in an immortalized mouse brain cell line (MBA 13). Intraperitoneal injection of 10 p.f.u. of CME2 into 4- to 6- week-old BALB/c mice caused mild clinical signs in some mice, whereas the majority of the infected animals remained asymptomatic, similar to infection with the avirulent SFV A7. In contrast, infection with the parental SFV4, a derivative of the virulent L10 strain, was lethal in 80% of mice. Virus titres in blood and brain tissue specimens of BALB/c mice were similar after infection with CME2 or A7 viruses. The results suggest that amino acid differences in the E2 glycoprotein individually or in concert cause the attenuation of CME2.

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1995-01-01
2022-10-04
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References

  1. Balluz I. M., Glasgow G. M., Killen H. M., Mabruk M. J. M.E. F., Sheahan B.J., Atkins G. J. 1993; Virulent and avirulent strains of Semliki Forest virus show similar cell tropism for the murine central nervous system but differ in the severity and rate of induction of cytolytic damage. Neuropathology and Applied Neurobiology 19:233–239
    [Google Scholar]
  2. Boere W. A. M., Benaissa-Trouw B. J., Harmsen M., Kraaijeveld C. A., Snippe H. 1983; Neutralizing and non-neutralizing monoclonal antibodies to the E2 glycoprotein of Semliki Forest virus can protect mice from lethal encephalitis. Journal of General Virology 64:1405–1408
    [Google Scholar]
  3. Bradish C. J., Allner K., Maber H. B. 1971; The virulence of original and derived strains of Semliki Forest virus for mice, guinea pigs and rabbits. Journal of General Virology 12:141–162
    [Google Scholar]
  4. Davis N. L., Powell N., Greenwald G. F., Willis F. V., Johnson B. J. B., Smith J. F., Johnston R. E. 1991; Attenuating mutations in the E2 glycoprotein gene of Venezuelan equine encephalitis virus: construction of single and multiple mutants in a full-length cDNA clone. Virology 183:20–31
    [Google Scholar]
  5. De Curtis I., Simons K. 1986; Dissection of Semliki Forest virus glycoprotein delivery from the trans-golgi network to the cell surface in permeabilized BHK cells. Proceedings of the National Academy of SciencesUSA 85:8052–8056
    [Google Scholar]
  6. Faragher S. G., Meek A. D., Rice C. M., Dalgarno L. 1988; Genome sequences of a mouse-avirulent and mouse-virulent strain of Ross River virus. Virology 163:509–526
    [Google Scholar]
  7. Fazakerley J. K., Webb H. E. 1987; Semliki Forest virus- induced, immune-mediated demyelination: adoptive transfer studies and viral persistence in nude mice. Journal of General Virology 68:377–385
    [Google Scholar]
  8. Fazakereley J. K., Pathak S., Scallan M., Amor S., Dyson H. 1993; Replication of the A7 (74) strain of Semliki Forest virus is restricted in neurons. Virology 195:627–637
    [Google Scholar]
  9. Garoff H. A., Frischauf A. -M., Simons K., Fehrach H., Delius H. 1980a; Nucleotide sequence of cDNA coding for Semliki Forest virus membrane glycoproteins. Nature 288:236–241
    [Google Scholar]
  10. Garoff H., Frischauf A. -M., Simons K., Fehrach H., Delius H. 1980b; The capsid protein of Semliki Forest virus has clusters of basic amino acids and prolines in its amino-terminal region. Proceedings of the National Academy of Sciences USA 77:6376–6380
    [Google Scholar]
  11. Gates M.C., Sheahan V. J., O’Sullivan M. A., Atkins G. J. 1985; The pathogenenicity of the A7, M9 and F10 strains of Semliki Forest virus for weanling mice and primary mouse brain cell cultures. Journal of General Virology 66:2365–2373
    [Google Scholar]
  12. Glasgow G. M., Sheahan B. J., Atkins G. J., Wahlberg J. M., Salminen A., Liljeström P. 1991; Two mutations in the envelope glycoprotein E2 of Semliki Forest virus affecting the maturation and entry patterns of the virus alter pathogenicity for mice. Virology 185:741–748
    [Google Scholar]
  13. Glasgow G. M., Killen H. M., Liljeström P., Sheahan B., Atkins G. J. 1994; A single amino acid change in the E2 spike protein of a virulent strain of Semliki Forest virus attenuates pathogenicity. Journal of General Virology 75:663–668
    [Google Scholar]
  14. Kinney R. M., Chang G.-J., Tsuchiya K. R., Sneider J. M., Roehrig J. T., Woodward T. M., Trent D. W. 1993; Attenuation of Venezuelan equine encephalitis virus strain TC-83 is encoded by the 5′-noncoding region and the E2 envelope gly coprotein. Journal of Virology 67:1269–1277
    [Google Scholar]
  15. Liljeström P., Lusa S., Huylebroeck D., Garoff H. 1991; In vitro mutagenesis of a full-length cDNA clone of Semliki Forest virus: the small 6,000-molecular-weight membrane protein modulates virus release. Journal of Virology 65:4107–4113
    [Google Scholar]
  16. Lustig S., Jackson A. C., Hahn C. S., Griffin D. E., Strauss E. G., Strauss J. H. 1988; Molecular basis of Sindbis virus neurovirulence in mice. Journal of Virology 62:2329–2336
    [Google Scholar]
  17. Melancon P., Garoff H. 1987; Processing of the Semliki Forest virus structural polyprotein: role of the capsid protease. Journal of Virology 61:1301–1309
    [Google Scholar]
  18. Ou J. -H., Strauss E. G., Strauss H. H. 1981; Comparative studies of the 3′ terminal sequences of several alphavirus RNAs. Virology 109:281–289
    [Google Scholar]
  19. Pence D. F., Davis N. L., Johnston R. E. 1990; Antigenic and genetic characterization of Sindbis virus monoclonal antibody escape mutants which define a pathogenesis domain on glycoprotein E2. Virology 175:41–49
    [Google Scholar]
  20. Polo J. M., Johnston R. E. 1990; Attenuating mutations in glycoproteins El and E2 of Sindbis virus produce a highly attenuated strain when combined in vitro. Journal of Virology 64:4438–1444
    [Google Scholar]
  21. Russel D. L., Dalrymple J. M., Johnston R. E. 1989; Sindbis virus mutations which coordinatively affect glycoprotein processing, penetration, and virulence in mice. Journal of Virology 63:1619–1629
    [Google Scholar]
  22. Sanger F., Niklen S., Coulson A. R. 1977; DNA sequencing with chain-termination inhibitors. Proceedings of the National Academy of SciencesUSA 74:5463–5467
    [Google Scholar]
  23. Santagati M. G., Itäranta P. V., Koskimies P. V., Määttä J. A., Salmi A. A., Hinkkanen A. E. 1994; Multiple repeating motifs are found in the 3′-terminal non-translated region of Semliki Forest virus A7 variant genome. Journal of General Virology 75:1499–1504
    [Google Scholar]
  24. Smyth J. M. B., Sheahan B. J., Atkins G. J. 1990; Multiplication of virulent and demyelinating Semliki Forest virus in the mouse central nervous system: consequences in BALB/c and SJL mice. Journal of General Virology 71:2575–2583
    [Google Scholar]
  25. Syväoja P., Peränen J., Suomalainen M., Keränen S., & Kääriäinen L. 1990; A single amino acid change in E3 of tsl mutant inhibits the intracellular transport of SFV envelope protein complex. Virology 179:658–666
    [Google Scholar]
  26. Takkinen K. 1986; Complete nucleotide sequence of the non- structural protein genes of Semliki Forest virus. Nucleic Acids Research 14:5667–5682
    [Google Scholar]
  27. Tucker P. C., Griffin D. E. 1991; Mechanism of altered Sindbis virus neurovirulence associated with a single-amino-acid change in the E2 glycoprotein. Journal of Virology 65:155–1557
    [Google Scholar]
  28. Wu L.-X., Mäkelä M. J., Royttä M., Salmi A. 1988; Effect of viral infection on experimental allergic encephalomyelitis in mice. Journal of Neuroimmunology 18:139–153
    [Google Scholar]
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