1887

Abstract

The foot-and-mouth disease virus (FMDV) Leader (L) protein is produced in two forms, Lab and Lb, differing only at their amino-termini, due to the use of separate initiation codons, usually 84 nt apart. It has been shown previously, and confirmed here, that precise deletion of the Lab coding sequence is lethal for the virus, whereas loss of the Lb coding sequence results in a virus that is viable in BHK cells. In addition, it is now shown that deletion of the ‘spacer’ region between these two initiation codons can be tolerated. Growth of the virus precisely lacking just the Lb coding sequence resulted in a previously undetected accumulation of frameshift mutations within the ‘spacer’ region. These mutations block the inappropriate fusion of amino acid sequences to the amino-terminus of the capsid protein precursor. Modification, by site-directed mutagenesis, of the Lab initiation codon, in the context of the virus lacking the Lb coding region, was also tolerated by the virus within BHK cells. However, precise loss of the Lb coding sequence alone blocked FMDV replication in primary bovine thyroid cells. Thus, the requirement for the Leader protein coding sequences is highly dependent on the nature and extent of the residual Leader protein sequences and on the host cell system used. FMDVs precisely lacking Lb and with the Lab initiation codon modified may represent safer seed viruses for vaccine production.

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2013-07-01
2019-12-08
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References

  1. Abrams C. C., King A. M. Q., Belsham G. J.. ( 1995; ). Assembly of foot-and-mouth disease virus empty capsids synthesized by a vaccinia virus expression system. . J Gen Virol 76:, 3089–3098. [CrossRef] [PubMed]
    [Google Scholar]
  2. Andreev D. E., Fernandez-Miragall O., Ramajo J., Dmitriev S. E., Terenin I. M., Martinez-Salas E., Shatsky I. N.. ( 2007; ). Differential factor requirement to assemble translation initiation complexes at the alternative start codons of foot-and-mouth disease virus RNA. . RNA 13:, 1366–1374. [CrossRef] [PubMed]
    [Google Scholar]
  3. Belsham G. J.. ( 1992; ). Dual initiation sites of protein synthesis on foot-and-mouth disease virus RNA are selected following internal entry and scanning of ribosomes in vivo, . EMBO J 11:, 1105–1110.[PubMed]
    [Google Scholar]
  4. Belsham G. J.. ( 2005; ). Translation and replication of FMDV RNA. . Curr Top Microbiol Immunol 288:, 43–70. [CrossRef] [PubMed]
    [Google Scholar]
  5. Belsham G. J., Brangwyn J. K.. ( 1990; ). A region of the 5′ noncoding region of foot-and-mouth disease virus RNA directs efficient internal initiation of protein synthesis within cells: involvement with the role of L protease in translational control. . J Virol 64:, 5389–5395.[PubMed]
    [Google Scholar]
  6. Belsham G. J., Abrams C. C., King A. M. Q., Roosien J., Vlak J. M.. ( 1991; ). Myristoylation of foot-and-mouth disease virus capsid protein precursors is independent of other viral proteins and occurs in both mammalian and insect cells. . J Gen Virol 72:, 747–751. [CrossRef] [PubMed]
    [Google Scholar]
  7. Belsham G. J., McInerney G. M., Ross-Smith N.. ( 2000; ). Foot-and-mouth disease virus 3C protease induces cleavage of translation initiation factors eIF4A and eIF4G within infected cells. . J Virol 74:, 272–280. [CrossRef] [PubMed]
    [Google Scholar]
  8. Belsham G. J., Nielsen I., Normann P., Royall E., Roberts L. O.. ( 2008; ). Monocistronic mRNAs containing defective hepatitis C virus-like picornavirus internal ribosome entry site elements in their 5′ untranslated regions are efficiently translated in cells by a cap-dependent mechanism. . RNA 14:, 1671–1680. [CrossRef] [PubMed]
    [Google Scholar]
  9. Bøtner A., Kakker N. K., Barbezange C., Berryman S., Jackson T., Belsham G. J.. ( 2011; ). Capsid proteins from field strains of foot-and-mouth disease virus confer a pathogenic phenotype in cattle on an attenuated, cell-culture-adapted virus. . J Gen Virol 92:, 1141–1151. [CrossRef] [PubMed]
    [Google Scholar]
  10. Brown C. C., Piccone M. E., Mason P. W., McKenna T. S., Grubman M. J.. ( 1996; ). Pathogenesis of wild-type and leaderless foot-and-mouth disease virus in cattle. . J Virol 70:, 5638–5641.[PubMed]
    [Google Scholar]
  11. Cao X., Bergmann I. E., Füllkrug R., Beck E.. ( 1995; ). Functional analysis of the two alternative translation initiation sites of foot-and-mouth disease virus. . J Virol 69:, 560–563.[PubMed]
    [Google Scholar]
  12. Carrillo C., Tulman E. R., Delhon G., Lu Z., Carreno A., Vagnozzi A., Kutish G. F., Rock D. L.. ( 2005; ). Comparative genomics of foot-and-mouth disease virus. . J Virol 79:, 6487–6504. [CrossRef] [PubMed]
    [Google Scholar]
  13. Chow M., Newman J. F., Filman D., Hogle J. M., Rowlands D. J., Brown F.. ( 1987; ). Myristylation of picornavirus capsid protein VP4 and its structural significance. . Nature 327:, 482–486. [CrossRef] [PubMed]
    [Google Scholar]
  14. de los Santos T., de Avila Botton S., Weiblen R., Grubman M. J.. ( 2006; ). The leader proteinase of foot-and-mouth disease virus inhibits the induction of beta interferon mRNA and blocks the host innate immune response. . J Virol 80:, 1906–1914. [CrossRef] [PubMed]
    [Google Scholar]
  15. de los Santos T., Diaz-San Segundo F., Grubman M. J.. ( 2007; ). Degradation of nuclear factor kappa B during foot-and-mouth disease virus infection. . J Virol 81:, 12803–12815. [CrossRef] [PubMed]
    [Google Scholar]
  16. Ellard F. M., Drew J., Blakemore W. E., Stuart D. I., King A. M. Q.. ( 1999; ). Evidence for the role of His-142 of protein 1C in the acid-induced disassembly of foot-and-mouth disease virus capsids. . J Gen Virol 80:, 1911–1918.[PubMed]
    [Google Scholar]
  17. Fuerst T. R., Niles E. G., Studier F. W., Moss B.. ( 1986; ). Eukaryotic transient-expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase. . Proc Natl Acad Sci U S A 83:, 8122–8126. [CrossRef] [PubMed]
    [Google Scholar]
  18. Gradi A., Foeger N., Strong R., Svitkin Y. V., Sonenberg N., Skern T., Belsham G. J.. ( 2004; ). Cleavage of eukaryotic translation initiation factor 4GII within foot-and-mouth disease virus-infected cells: identification of the L-protease cleavage site in vitro. . J Virol 78:, 3271–3278. [CrossRef] [PubMed]
    [Google Scholar]
  19. Kaminski A., Howell M. T., Jackson R. J.. ( 1990; ). Initiation of encephalomyocarditis virus RNA translation: the authentic initiation site is not selected by a scanning mechanism. . EMBO J 9:, 3753–3759.[PubMed]
    [Google Scholar]
  20. Kaminski A., Belsham G. J., Jackson R. J.. ( 1994; ). Translation of encephalomyocarditis virus RNA: parameters influencing the selection of the internal initiation site. . EMBO J 13:, 1673–1681.[PubMed]
    [Google Scholar]
  21. Kirchweger R., Ziegler E., Lamphear B. J., Waters D., Liebig H. D., Sommergruber W., Sobrino F., Hohenadl C., Blaas D.. & other authors ( 1994; ). Foot-and-mouth disease virus leader proteinase: purification of the Lb form and determination of its cleavage site on eIF-4 gamma. . J Virol 68:, 5677–5684.[PubMed]
    [Google Scholar]
  22. Kong W. P., Roos R. P.. ( 1991; ). Alternative translation initiation site in the DA strain of Theiler’s murine encephalomyelitis virus. . J Virol 65:, 3395–3399.[PubMed]
    [Google Scholar]
  23. Lohse L., Jackson T., Bøtner A., Belsham G. J.. ( 2012; ). Capsid coding sequences of foot-and-mouth disease viruses are determinants of pathogenicity in pigs. . Vet Res 43:, 46. [CrossRef] [PubMed]
    [Google Scholar]
  24. López de Quinto S., Martínez-Salas E.. ( 1999; ). Involvement of the aphthovirus RNA region located between the two functional AUGs in start codon selection. . Virology 255:, 324–336. [CrossRef] [PubMed]
    [Google Scholar]
  25. Loughran G., Firth A. E., Atkins J. F.. ( 2011; ). Ribosomal frameshifting into an overlapping gene in the 2B-encoding region of the cardiovirus genome. . Proc Natl Acad Sci U S A 108:, E1111–E1119. [CrossRef] [PubMed]
    [Google Scholar]
  26. Medina M., Domingo E., Brangwyn J. K., Belsham G. J.. ( 1993; ). The two species of the foot-and-mouth disease virus leader protein, expressed individually, exhibit the same activities. . Virology 194:, 355–359. [CrossRef] [PubMed]
    [Google Scholar]
  27. Nayak A., Goodfellow I. G., Woolaway K. E., Birtley J., Curry S., Belsham G. J.. ( 2006; ). Role of RNA structure and RNA binding activity of foot-and-mouth disease virus 3C protein in VPg uridylylation and virus replication. . J Virol 80:, 9865–9875. [CrossRef] [PubMed]
    [Google Scholar]
  28. Ohlmann T., Jackson R. J.. ( 1999; ). The properties of chimeric picornavirus IRESes show that discrimination between internal translation initiation sites is influenced by the identity of the IRES and not just the context of the AUG codon. . RNA 5:, 764–778. [CrossRef] [PubMed]
    [Google Scholar]
  29. Piccone M. E., Rieder E., Mason P. W., Grubman M. J.. ( 1995; ). The foot-and-mouth disease virus leader proteinase gene is not required for viral replication. . J Virol 69:, 5376–5382.[PubMed]
    [Google Scholar]
  30. Piccone M. E., Pacheco J. M., Pauszek S. J., Kramer E., Rieder E., Borca M. V., Rodriguez L. L.. ( 2010; ). The region between the two polyprotein initiation codons of foot-and-mouth disease virus is critical for virulence in cattle. . Virology 396:, 152–159. [CrossRef] [PubMed]
    [Google Scholar]
  31. Piccone M. E., Diaz-San Segundo F., Kramer E., Rodriguez L. L., de los Santos T.. ( 2011; ). Introduction of tag epitopes in the inter-AUG region of foot and mouth disease virus: effect on the L protein. . Virus Res 155:, 91–97. [CrossRef] [PubMed]
    [Google Scholar]
  32. Polacek C., Gullberg M., Li J., Belsham G. J.. ( 2013; ) Low levels of foot-and-mouth disease virus 3Cpro expression are required to achieve optimal capsid protein expression and processing in mammalian cells. . J Gen Virol vir.0.050492-0; published ahead of print January 30, 2013 . [CrossRef]
    [Google Scholar]
  33. Pöyry T. A., Hentze M. W., Jackson R. J.. ( 2001; ). Construction of regulatable picornavirus IRESes as a test of current models of the mechanism of internal translation initiation. . RNA 7:, 647–660. [CrossRef] [PubMed]
    [Google Scholar]
  34. Reed L. J., Muench H.. ( 1938; ). A simple method of estimating fifty percent endpoints. . Am J Hyg 27:, 493–497.
    [Google Scholar]
  35. Reid S. M., Grierson S. S., Ferris N. P., Hutchings G. H., Alexandersen S.. ( 2003; ). Evaluation of automated RT-PCR to accelerate the laboratory diagnosis of foot-and-mouth disease virus. . J Virol Methods 107:, 129–139. [CrossRef] [PubMed]
    [Google Scholar]
  36. Roeder P. L., Le Blanc Smith P. M.. ( 1987; ). Detection and typing of foot-and-mouth disease virus by enzyme-linked immunosorbent assay: a sensitive, rapid and reliable technique for primary diagnosis. . Res Vet Sci 43:, 225–232.[PubMed]
    [Google Scholar]
  37. Sangar D. V., Newton S. E., Rowlands D. J., Clarke B. E.. ( 1987; ). All foot and mouth disease virus serotypes initiate protein synthesis at two separate AUGs. . Nucleic Acids Res 15:, 3305–3315. [CrossRef] [PubMed]
    [Google Scholar]
  38. Strong R., Belsham G. J.. ( 2004; ). Sequential modification of translation initiation factor eIF4GI by two different foot-and-mouth disease virus proteases within infected baby hamster kidney cells: identification of the 3Cpro cleavage site. . J Gen Virol 85:, 2953–2962. [CrossRef] [PubMed]
    [Google Scholar]
  39. Towler D. A., Gordon J. I., Adams S. P., Glaser L.. ( 1988; ). The biology and enzymology of eukaryotic protein acylation. . Annu Rev Biochem 57:, 69–97. [CrossRef] [PubMed]
    [Google Scholar]
  40. Uddowla S., Hollister J., Pacheco J. M., Rodriguez L. L., Rieder E.. ( 2012; ). A safe foot-and-mouth disease vaccine platform with two negative markers for differentiating infected from vaccinated animals. . J Virol 86:, 11675–11685. [CrossRef] [PubMed]
    [Google Scholar]
  41. van Eyll O., Michiels T.. ( 2002; ). Non-AUG-initiated internal translation of the L* protein of Theiler’s virus and importance of this protein for viral persistence. . J Virol 76:, 10665–10673. [CrossRef] [PubMed]
    [Google Scholar]
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