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. 1995 Feb;69(2):864–870. doi: 10.1128/jvi.69.2.864-870.1995

Attenuation of Theiler's murine encephalomyelitis virus by modifications of the oligopyrimidine/AUG tandem, a host-dependent translational cis element.

E V Pilipenko 1, A P Gmyl 1, S V Maslova 1, E V Khitrina 1, V I Agol 1
PMCID: PMC188653  PMID: 7815554

Abstract

A set of Theiler's murine encephalomyelitis virus mutants with engineered alterations in the conserved oligopyrimidine/AUG tandem (E. V. Pilipenko, A. P. Gmyl, S. V. Maslova, G. A. Belov, A. N. Sinyakov, M. Huang, T. D. K. Brown, and V. I. Agol, J. Mol. Biol. 241:398-414, 1994) were assayed for their growth potential in BHK-21 cells (as reflected in plaque size) and for neurovirulence upon intracerebral inoculation of mice. Tandem-destroying mutations, which included substitutions in the oligopyrimidine moiety and extended insertions into the oligopyrimidine/AUG spacer, exerted relatively little effect on the plaque size but ensured a high level of attenuation. The attenuated mutants exhibited remarkable genetic stability upon growth in BHK-21 cells. However, the brains of rare animals that developed symptoms after the inoculation with high doses of these mutants invariably contained pseudorevertants with the oligopyrimidine/AUG tandem restored by diverse deletions or an AUG-generating point mutation. The AUG moiety of the tandem in the revertant genomes was represented by either a cryptic codon or initiator codon. The results demonstrate that the tandem, while dispensable for the Theiler's murine encephalomyelitis virus growth in BHK-21 cells, is essential for neurovirulence in mice. Thus, the oligopyrimidine/AUG tandem is a host-dependent cis-acting control element that may be essential for virus replication under certain conditions. The functional activity of the tandem was retained when its oligopyrimidine or AUG moieties were made double stranded. A possible role of the tandem in the cap-independent internal initiation of translation on the picornavirus RNA templates is discussed.

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Selected References

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  1. Agol V. I. The 5'-untranslated region of picornaviral genomes. Adv Virus Res. 1991;40:103–180. doi: 10.1016/S0065-3527(08)60278-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bandyopadhyay P. K., Pritchard A., Jensen K., Lipton H. L. A three-nucleotide insertion in the H stem-loop of the 5' untranslated region of Theiler's virus attenuates neurovirulence. J Virol. 1993 Jun;67(6):3691–3695. doi: 10.1128/jvi.67.6.3691-3695.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beck E., Forss S., Strebel K., Cattaneo R., Feil G. Structure of the FMDV translation initiation site and of the structural proteins. Nucleic Acids Res. 1983 Nov 25;11(22):7873–7885. doi: 10.1093/nar/11.22.7873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gmyl A. P., Pilipenko E. V., Maslova S. V., Belov G. A., Agol V. I. Functional and genetic plasticities of the poliovirus genome: quasi-infectious RNAs modified in the 5'-untranslated region yield a variety of pseudorevertants. J Virol. 1993 Oct;67(10):6309–6316. doi: 10.1128/jvi.67.10.6309-6316.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Iizuka N., Kohara M., Hagino-Yamagishi K., Abe S., Komatsu T., Tago K., Arita M., Nomoto A. Construction of less neurovirulent polioviruses by introducing deletions into the 5' noncoding sequence of the genome. J Virol. 1989 Dec;63(12):5354–5363. doi: 10.1128/jvi.63.12.5354-5363.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Jang S. K., Pestova T. V., Hellen C. U., Witherell G. W., Wimmer E. Cap-independent translation of picornavirus RNAs: structure and function of the internal ribosomal entry site. Enzyme. 1990;44(1-4):292–309. doi: 10.1159/000468766. [DOI] [PubMed] [Google Scholar]
  7. Jarousse N., Fiette L., Grant R. A., Hogle J. M., McAllister A., Michiels T., Aubert C., Tangy F., Brahic M., Peña Rossi C. Chimeric Theiler's virus with altered tropism for the central nervous system. J Virol. 1994 May;68(5):2781–2786. doi: 10.1128/jvi.68.5.2781-2786.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jarousse N., Grant R. A., Hogle J. M., Zhang L., Senkowski A., Roos R. P., Michiels T., Brahic M., McAllister A. A single amino acid change determines persistence of a chimeric Theiler's virus. J Virol. 1994 May;68(5):3364–3368. doi: 10.1128/jvi.68.5.3364-3368.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kaminski A., Belsham G. J., Jackson R. J. Translation of encephalomyocarditis virus RNA: parameters influencing the selection of the internal initiation site. EMBO J. 1994 Apr 1;13(7):1673–1681. doi: 10.1002/j.1460-2075.1994.tb06431.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kühn R., Luz N., Beck E. Functional analysis of the internal translation initiation site of foot-and-mouth disease virus. J Virol. 1990 Oct;64(10):4625–4631. doi: 10.1128/jvi.64.10.4625-4631.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. La Monica N., Racaniello V. R. Differences in replication of attenuated and neurovirulent polioviruses in human neuroblastoma cell line SH-SY5Y. J Virol. 1989 May;63(5):2357–2360. doi: 10.1128/jvi.63.5.2357-2360.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Meerovitch K., Nicholson R., Sonenberg N. In vitro mutational analysis of cis-acting RNA translational elements within the poliovirus type 2 5' untranslated region. J Virol. 1991 Nov;65(11):5895–5901. doi: 10.1128/jvi.65.11.5895-5901.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Nicholson R., Pelletier J., Le S. Y., Sonenberg N. Structural and functional analysis of the ribosome landing pad of poliovirus type 2: in vivo translation studies. J Virol. 1991 Nov;65(11):5886–5894. doi: 10.1128/jvi.65.11.5886-5894.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pestova T. V., Hellen C. U., Wimmer E. Translation of poliovirus RNA: role of an essential cis-acting oligopyrimidine element within the 5' nontranslated region and involvement of a cellular 57-kilodalton protein. J Virol. 1991 Nov;65(11):6194–6204. doi: 10.1128/jvi.65.11.6194-6204.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pilipenko E. V., Blinov V. M., Chernov B. K., Dmitrieva T. M., Agol V. I. Conservation of the secondary structure elements of the 5'-untranslated region of cardio- and aphthovirus RNAs. Nucleic Acids Res. 1989 Jul 25;17(14):5701–5711. doi: 10.1093/nar/17.14.5701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pilipenko E. V., Blinov V. M., Romanova L. I., Sinyakov A. N., Maslova S. V., Agol V. I. Conserved structural domains in the 5'-untranslated region of picornaviral genomes: an analysis of the segment controlling translation and neurovirulence. Virology. 1989 Feb;168(2):201–209. doi: 10.1016/0042-6822(89)90259-6. [DOI] [PubMed] [Google Scholar]
  17. Pilipenko E. V., Gmyl A. P., Maslova S. V., Belov G. A., Sinyakov A. N., Huang M., Brown T. D., Agol V. I. Starting window, a distinct element in the cap-independent internal initiation of translation on picornaviral RNA. J Mol Biol. 1994 Aug 19;241(3):398–414. doi: 10.1006/jmbi.1994.1516. [DOI] [PubMed] [Google Scholar]
  18. Pilipenko E. V., Gmyl A. P., Maslova S. V., Svitkin Y. V., Sinyakov A. N., Agol V. I. Prokaryotic-like cis elements in the cap-independent internal initiation of translation on picornavirus RNA. Cell. 1992 Jan 10;68(1):119–131. doi: 10.1016/0092-8674(92)90211-t. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Sonenberg N., Meerovitch K. Translation of poliovirus mRNA. Enzyme. 1990;44(1-4):278–291. doi: 10.1159/000468765. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Svitkin Y. V., Cammack N., Minor P. D., Almond J. W. Translation deficiency of the Sabin type 3 poliovirus genome: association with an attenuating mutation C472----U. Virology. 1990 Mar;175(1):103–109. doi: 10.1016/0042-6822(90)90190-3. [DOI] [PubMed] [Google Scholar]
  23. Svitkin Y. V., Maslova S. V., Agol V. I. The genomes of attenuated and virulent poliovirus strains differ in their in vitro translation efficiencies. Virology. 1985 Dec;147(2):243–252. doi: 10.1016/0042-6822(85)90127-8. [DOI] [PubMed] [Google Scholar]
  24. Svitkin Y. V., Pestova T. V., Maslova S. V., Agol V. I. Point mutations modify the response of poliovirus RNA to a translation initiation factor: a comparison of neurovirulent and attenuated strains. Virology. 1988 Oct;166(2):394–404. doi: 10.1016/0042-6822(88)90510-7. [DOI] [PubMed] [Google Scholar]
  25. Wimmer E., Hellen C. U., Cao X. Genetics of poliovirus. Annu Rev Genet. 1993;27:353–436. doi: 10.1146/annurev.ge.27.120193.002033. [DOI] [PubMed] [Google Scholar]
  26. Zurbriggen A., Thomas C., Yamada M., Roos R. P., Fujinami R. S. Direct evidence of a role for amino acid 101 of VP-1 in central nervous system disease in Theiler's murine encephalomyelitis virus infection. J Virol. 1991 Apr;65(4):1929–1937. doi: 10.1128/jvi.65.4.1929-1937.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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