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. 1993 Jul;67(7):3901–3907. doi: 10.1128/jvi.67.7.3901-3907.1993

Assembly of Theiler's virus recombinants used in mapping determinants of neurovirulence.

A E Pritchard 1, K Jensen 1, H L Lipton 1
PMCID: PMC237756  PMID: 8510210

Abstract

A major determinant of neurovirulence for the GDVII strain of Theiler's virus, a murine picornavirus, was mapped to the P1 capsid protein region. Chimeric viruses were constructed by using sequences from the 5' noncoding and P1 regions of the virulent GDVII strain to replace equivalent regions of the less virulent BeAn strain. Neurovirulence in mice progressively increased as larger regions of BeAn capsid protein-encoding sequences were replaced. The in vitro growth characteristics of the chimeras showed that some chimeras were growth delayed in BHK-21 cells even though the viral constructs exhibited larger plaque sizes, were less temperature sensitive, and were more thermally stable than BeAn. Examination of assembly intermediates revealed an altered pentamer conformation and delayed empty capsid formation for the growth-compromised viruses. For these constructs, their chimeric nature inadvertently resulted in virion assembly defects that complicated finer-scale mapping of the determinants of virulence within the capsid region. These results demonstrate the importance of determining in vitro growth characteristics of chimeras to correctly decipher the significance of their phenotypes. VP1 does not contain a complete determinate for virulence because a chimera with VP1-encoding sequences from GDVII in an otherwise BeAn virus has an attenuated phenotype but is not growth compromised in vitro. The source of sequences, BeAn or GDVII, in the 5' noncoding region had only slight effects on the virulence of recombinant constructs.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Calenoff M. A., Faaberg K. S., Lipton H. L. Genomic regions of neurovirulence and attenuation in Theiler murine encephalomyelitis virus. Proc Natl Acad Sci U S A. 1990 Feb;87(3):978–982. doi: 10.1073/pnas.87.3.978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Fu J. L., Stein S., Rosenstein L., Bodwell T., Routbort M., Semler B. L., Roos R. P. Neurovirulence determinants of genetically engineered Theiler viruses. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4125–4129. doi: 10.1073/pnas.87.11.4125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Fu J., Rodriguez M., Roos R. P. Strains from both Theiler's virus subgroups encode a determinant for demyelination. J Virol. 1990 Dec;64(12):6345–6348. doi: 10.1128/jvi.64.12.6345-6348.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Grant R. A., Filman D. J., Fujinami R. S., Icenogle J. P., Hogle J. M. Three-dimensional structure of Theiler virus. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2061–2065. doi: 10.1073/pnas.89.6.2061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kunkel T. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. doi: 10.1073/pnas.82.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  7. Lipton H. L., Calenoff M., Bandyopadhyay P., Miller S. D., Dal Canto M. C., Gerety S., Jensen K. The 5' noncoding sequences from a less virulent Theiler's virus dramatically attenuate GDVII neurovirulence. J Virol. 1991 Aug;65(8):4370–4377. doi: 10.1128/jvi.65.8.4370-4377.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lipton H. L. Persistent Theiler's murine encephalomyelitis virus infection in mice depends on plaque size. J Gen Virol. 1980 Jan;46(1):169–177. doi: 10.1099/0022-1317-46-1-169. [DOI] [PubMed] [Google Scholar]
  9. Lipton H. L. Theiler's virus infection in mice: an unusual biphasic disease process leading to demyelination. Infect Immun. 1975 May;11(5):1147–1155. doi: 10.1128/iai.11.5.1147-1155.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Luo M., He C., Toth K. S., Zhang C. X., Lipton H. L. Three-dimensional structure of Theiler murine encephalomyelitis virus (BeAn strain). Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2409–2413. doi: 10.1073/pnas.89.6.2409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Macadam A. J., Ferguson G., Arnold C., Minor P. D. An assembly defect as a result of an attenuating mutation in the capsid proteins of the poliovirus type 3 vaccine strain. J Virol. 1991 Oct;65(10):5225–5231. doi: 10.1128/jvi.65.10.5225-5231.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. McAllister A., Tangy F., Aubert C., Brahic M. Genetic mapping of the ability of Theiler's virus to persist and demyelinate. J Virol. 1990 Sep;64(9):4252–4257. doi: 10.1128/jvi.64.9.4252-4257.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Pritchard A. E., Calenoff M. A., Simpson S., Jensen K., Lipton H. L. A single base deletion in the 5' noncoding region of Theiler's virus attenuates neurovirulence. J Virol. 1992 Apr;66(4):1951–1958. doi: 10.1128/jvi.66.4.1951-1958.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Stein S. B., Zhang L., Roos R. P. Influence of Theiler's murine encephalomyelitis virus 5' untranslated region on translation and neurovirulence. J Virol. 1992 Jul;66(7):4508–4517. doi: 10.1128/jvi.66.7.4508-4517.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Tangy F., McAllister A., Aubert C., Brahic M. Determinants of persistence and demyelination of the DA strain of Theiler's virus are found only in the VP1 gene. J Virol. 1991 Mar;65(3):1616–1618. doi: 10.1128/jvi.65.3.1616-1618.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Zurbriggen A., Hogle J. M., Fujinami R. S. Alteration of amino acid 101 within capsid protein VP-1 changes the pathogenicity of Theiler's murine encephalomyelitis virus. J Exp Med. 1989 Dec 1;170(6):2037–2049. doi: 10.1084/jem.170.6.2037. [DOI] [PMC free article] [PubMed] [Google Scholar]

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