1887

Abstract

The unique region of murine hepatitis virus (MHV) mRNA 5 has two open reading frames, ORF 5a and ORF 5b, that encode small proteins of unknown function. In the experiments described here, we have used the translation of synthetic mRNAs to examine the expression of these ORFs. Our results show that a synthetic mRNA containing both ORFs is functionally bicistronic. More importantly, the expression of ORF 5b, but not ORF 5a, is maintained in a trieistronic mRNA containing an additional 5′-proximal ORF. Thus, in the context of the MHV mRNA 5 unique region, the initiation of protein synthesis on ORF 5b can occur independently of ribosomes that enter from the 5′ end of the mRNA. We conclude that the translation of ORF 5b is mediated by the internal entry of ribosomes.

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1994-11-01
2022-01-17
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References

  1. Baker S. C., Shieh C. K., Soe L. H., Chang M. F., Vannier D. M., Lai M. M. C. 1989; Identification of a domain required for autoproteolytic cleavage of murine coronavirus gene A polyprotein. Journal of Virology 63:3693–3699
    [Google Scholar]
  2. Bonilla P. J., Gorbalenya A. E., Weiss S. R. 1994; Mouse hepatitis virus strain A59 polymerase gene ORF la: heterogeneity among MHV strains. Virology 198:736–740
    [Google Scholar]
  3. Bredenbeek P. J., Pachuk C. J., Noten A. F., Charite J., Luytjes W., Weiss S. R., Spaan W. J. 1990; The primary structure and expression of the second open reading frame of the polymerase gene of the coronavirus MHV-A59; a highly conserved polymerase is expressed by an efficient ribosomal frameshifting mechanism. Nucleic Acids Research 18:1825–1832
    [Google Scholar]
  4. Brown E. A., Zhang H., Ping L., Lemon S. M. 1992; Secondary structure of the 5′ nontranslated regions of hepatitis C virus and pestivirus genomic RNAs. Nucleic Acids Research 20:5041–5045
    [Google Scholar]
  5. Budzilowicz C. J., Weiss S. R. 1987; In vitro synthesis of two polypeptides from a nonstructural gene of coronavirus mouse hepatitis virus strain A59. Virology 157:509–515
    [Google Scholar]
  6. Cavanagh D., Brian D. A., Brinton M., Enjuanes L., Holmes K. V., Horzinek M.C, Laude H., Plagemann P. G., Siddell S., Spaan W. J. M., Taguchi F., Talbot P. J. 1994; Revision of the taxonomy of the Coronavirus, Torovirus and Arterivirus genera. Archives of Virology 135:227–236
    [Google Scholar]
  7. Contreras R., Cheroutre H., Degrave W., Fiers W. 1982; Simple efficient in vitro synthesis of capped RNA useful for direct expression of cloned DNA. Nucleic Acids Research 10:6353–6362
    [Google Scholar]
  8. Denison M. R., Zoltick P. W., Leibowitz J. L., Pachuk C. J., Weiss J. R. 1991; Identification of polypeptides encoded in open reading frame lb of the putative polymerase gene of murine coronavirus mouse hepatitis virus A59. Journal of Virology 65:3076–3082
    [Google Scholar]
  9. Denison M. R., Zoltick P. W., Hughes S. A., Giangreco B., Olson A. L., Perlman S., Leibowitz J. L., Weiss S. R. 1992; Intracellular processing of the N-terminal ORF la proteins of the coronavirus MHV A59 requires multiple proteolytic events. Virology 189:274–284
    [Google Scholar]
  10. Ebner D., Raabe T., Siddell S. G. 1988; Identification of the coronavirus MHV-JHM mRNA 4 product. Journal of General Virology 69:1041–1050
    [Google Scholar]
  11. Ellinger S., Glockshuber R., Jahn G., Plückthun A. 1989; Cleavage of procaryotically expressed human immunodeficiency virus fusion proteins by factor Xa and application in Western blot (immunoblot) assays. Journal of Clinical Microbiology 27:971–976
    [Google Scholar]
  12. Godet M., L’Haridon R., Vautherot J. F., Laude H. 1992; TGEV corona virus ORF4 encodes a membrane protein that is incorporated into virions. Virology 188:666–675
    [Google Scholar]
  13. Jackson R. J., Howell M. T., Kaminsky A. 1990; The novel mechanism of initiation of picornavirus RNA translation. Trends in Biochemical Science 15:477–483
    [Google Scholar]
  14. Jang S. K., Wimmer E. 1990; Cap-independent translation of encephalomyocarditis virus RNA: structural elements of the internal ribosomal entry site and involvement of a cellular 57-kD RNA- binding protein. Genes and Development 4:1560–1572
    [Google Scholar]
  15. Kozak M. 1989; The scanning model for translation: an update. Journal of Cell Biology 108:229–241
    [Google Scholar]
  16. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  17. Lae M. M. C., Patton C. D., Baric R. S., Stohlman S. A. 1983; Presence of leader sequences in the mRNA of mouse hepatitis virus. Journal of Virology 46:1027–1033
    [Google Scholar]
  18. Le S. -Y., Chen J. -H., Sonenberg N., Maizel J. V.Jr 1992; Conserved tertiary structure elements in the 5′ untranslated region of human enteroviruses and rhinoviruses. Virology 191:858–866
    [Google Scholar]
  19. Le S. -Y., Chen J. -H., Sonenberg N., Maizel J. V.Jr 1993; Conserved tertiary structural elements in the 5′ nontranslated region of cardiovirus, apthovirus and hepatitis A virus RNAs. Nucleic Acids Research 21:2445–2451
    [Google Scholar]
  20. Le S. -Y., Sonenberg N., Maizel J. V.Jr 1994; Distinct structural elements and internal entry of ribosomes in mRNA 3 encoded by infectious bronchitis virus. Virology 198:405–411
    [Google Scholar]
  21. Lee H. J., Shieh C. K., Gorbalenya A. E., Koonin E. V., La M. N., Tuler J., Bagdzhadzhyan A., Lai M. M. C. 1991; The complete sequence (22 kilobases) of murine coronavirus gene 1 encoding the putative proteases and RNA polymerase. Virology 180:567–582
    [Google Scholar]
  22. Leibowitz J. L., Weiss S. R., Paavola E., Bond C. W. 1982; Cell-free translation of murine coronavirus RNA. Journal of Virology 43:905–913
    [Google Scholar]
  23. Leibowitz J. L., Perlman S., Weinstock G., Devries J. R., Budzilowicz C., Weissemann J. M., Weiss S. R. 1988; Detection of a murine coronavirus nonstructural protein encoded in a downstream open reading frame. Virology 164:156–164
    [Google Scholar]
  24. Liu D. X., Inglis S. C. 1991; Association of the infectious bronchitis virus 3c protein with the virion envelope. Virology 185:911–917
    [Google Scholar]
  25. Liu D. X., Inglis S. C. 1992; Internal entry of ribosomes on a tricistronic mRNA encoded by infectious bronchitis virus. Journal of Virology 66:6143–6154
    [Google Scholar]
  26. Meerovitch K., Sonenberg N. 1993; Internal initiation of picornavirus RNA translation. Seminars in Virology 4:217–227
    [Google Scholar]
  27. Meerovitch K., Svitkin Y. V., Lee H. S., Lejbkowicz F., Kenan D. J., Chan E. K. L., Agol V. I., Keene J. D., Sonenberg N. 1993; La autoantigen enhances and corrects aberrant translation of poliovirus RNA in reticulocyte lysate. Journal of Virology 67:3798–3807
    [Google Scholar]
  28. Melton D. A., Krieg P. A., Rabaghiati M. R., Maniatis T., Zinn K., Green M. D. 1984; Efficient in vitro synthesis of biologically active RNA and RNA hybridisation probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Research 12:7035–7056
    [Google Scholar]
  29. Pachuk C. J., Bredenbeek P. J., Zoltik P. W., Spaan W. J. M., Weiss S. R. 1989; Molecular cloning of the gene encoding the putative polymerase of mouse hepatitis coronavirus A59. Virology 171:141–148
    [Google Scholar]
  30. Pestova T. V., Hellen C. U. T., Wimmer E. 1991; 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. Journal of Virology 65:6194–6204
    [Google Scholar]
  31. Schreier M. H., Staehelin T. 1973; Initiation of mammalian protein synthesis: the importance of ribosome and initiation factor quality for the efficiency of in vitro systems. Journal of Molecular Biology 73:329–349
    [Google Scholar]
  32. Siddell S. 1983; Coronavirus JHM: coding assignments of subgenomic mRNAs. Journal of General Virology 64:113–125
    [Google Scholar]
  33. Skinner M. A., Ebner D., Siddell S. G. 1985; Coronavirus MHV-JHM mRNA 5 has a sequence arrangement which potentially allows translation of a second, downstream open reading frame. Journal of General Virology 66:581–592
    [Google Scholar]
  34. Yao Z., Jones D. H., Grose C. 1992; Site-directed mutagenesis of herpesvirus glycoprotein phosphorylation sites by recombination polymerase chain reaction. PCR Methods and Applications 1:205–207
    [Google Scholar]
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