An analysis by metabolic labelling of the encephalomyocarditis virus ribosomal frameshifting efficiency and stimulators Open Access

Abstract

Programmed −1 ribosomal frameshifting is a mechanism of gene expression whereby specific signals within messenger RNAs direct a proportion of ribosomes to shift −1 nt and continue translating in the new reading frame. Such frameshifting normally depends on an RNA structure stimulator 3′-adjacent to a ‘slippery’ heptanucleotide shift site sequence. Recently we identified an unusual frameshifting mechanism in encephalomyocarditis virus, where the stimulator involves a -acting virus protein. Thus, in contrast to other examples of −1 frameshifting, the efficiency of frameshifting in encephalomyocarditis virus is best studied in the context of virus infection. Here we use metabolic labelling to analyse the frameshifting efficiency of wild-type and mutant viruses. Confirming previous results, frameshifting depends on a G_GUU_UUU shift site sequence and a 3′-adjacent stem-loop structure, but is not appreciably affected by the ‘StopGo’ sequence present ~30 nt upstream. At late timepoints, frameshifting was estimated to be 46–76 % efficient.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.000888
2017-08-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/98/8/2100.html?itemId=/content/journal/jgv/10.1099/jgv.0.000888&mimeType=html&fmt=ahah

References

  1. Atkins JF, Loughran G, Bhatt PR, Firth AE, Baranov PV. Ribosomal frameshifting and transcriptional slippage: from genetic steganography and cryptography to adventitious use. Nucleic Acids Res 2016; 44:7007–7078 [View Article]
    [Google Scholar]
  2. Firth AE, Brierley I. Non-canonical translation in RNA viruses. J Gen Virol 2012; 93:1385–1409 [View Article][PubMed]
    [Google Scholar]
  3. Ryan MD, Drew J. Foot-and-mouth disease virus 2A oligopeptide mediated cleavage of an artificial polyprotein. EMBO J 1994; 13:928–933[PubMed]
    [Google Scholar]
  4. Hahn H, Palmenberg AC. Mutational analysis of the encephalomyocarditis virus primary cleavage. J Virol 1996; 70:6870–6875[PubMed]
    [Google Scholar]
  5. Loughran G, Firth AE, Atkins JF. Ribosomal frameshifting into an overlapping gene in the 2B-encoding region of the cardiovirus genome. Proc Natl Acad Sci USA 2011; 108:E1111E1119 [View Article][PubMed]
    [Google Scholar]
  6. Finch LK, Ling R, Napthine S, Olspert A, Michiels T et al. Characterization of ribosomal frameshifting in Theiler's Murine Encephalomyelitis Virus. J Virol 2015; 89:8580–8589 [View Article][PubMed]
    [Google Scholar]
  7. Napthine S, Ling R, Finch LK, Jones JD, Bell S et al. Protein-directed ribosomal frameshifting temporally regulates gene expression. Nat Commun 2017; 8:15582 [View Article][PubMed]
    [Google Scholar]
  8. Duke GM, Osorio JE, Palmenberg AC. Attenuation of Mengo virus through genetic engineering of the 5' noncoding poly(C) tract. Nature 1990; 343:474–476 [View Article][PubMed]
    [Google Scholar]
  9. Loughran G, Libbey JE, Uddowla S, Scallan MF, Ryan MD et al. Theiler's murine encephalomyelitis virus contrasts with encephalomyocarditis and foot-and-mouth disease viruses in its functional utilization of the StopGo non-standard translation mechanism. J Gen Virol 2013; 94:348–353 [View Article][PubMed]
    [Google Scholar]
  10. Aminev AG, Amineva SP, Palmenberg AC. Encephalomyocarditis viral protein 2A localizes to nucleoli and inhibits cap-dependent mRNA translation. Virus Res 2003; 95:45–57 [View Article][PubMed]
    [Google Scholar]
  11. Groppo R, Brown BA, Palmenberg AC. Mutational analysis of the EMCV 2A protein identifies a nuclear localization signal and an eIF4E binding site. Virology 2011; 410:257–267 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.000888
Loading
/content/journal/jgv/10.1099/jgv.0.000888
Loading

Data & Media loading...

Most cited Most Cited RSS feed