The small genome segment of Bunyamwera orthobunyavirus harbours a single transcription-termination signal Open Access

Abstract

Transcription termination of the mRNA produced from the small (S) genome segment of Bunyamwera orthobunyavirus (BUNV) has previously been mapped to two -acting sequences located within the 5′ UTR using a virus-free replication assay. The ability of these sequences to terminate transcription was attributed to the shared pentanucleotide motif 3′-UGUCG-5′. Taking advantage of our plasmid-based rescue system to generate recombinant viruses, we re-evaluated the importance of both pentanucleotide motifs as well as that of two other conserved sequences in transcription termination . Analysis of the 3′ ends of positive-stranded viral RNAs derived from the S segment revealed that only the region around the upstream pentanucleotide motif mediated transcription termination in cells infected with wild-type BUNV, leading to mRNAs that were about 100 nt shorter than antigenome RNA. Furthermore, the downstream motif was not recognized in recombinant viruses in which the upstream signal has been disrupted. Our results suggest that in the context of virus infection transcription termination of the BUNV S genome segment mRNA is exclusively directed by the upstream-termination signal. Interestingly, within this region we identified a motif similar to a transcription-termination sequence used by Rift Valley fever phlebovirus.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.042390-0
2012-07-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/93/7/1449.html?itemId=/content/journal/jgv/10.1099/vir.0.042390-0&mimeType=html&fmt=ahah

References

  1. Abraham G., Pattnaik A. K. 1983; Early RNA synthesis in Bunyamwera virus-infected cells. J Gen Virol 64:1277–1290 [View Article][PubMed]
    [Google Scholar]
  2. Albariño C. G., Bird B. H., Nichol S. T. 2007; A shared transcription termination signal on negative and ambisense RNA genome segments of Rift Valley fever, sandfly fever Sicilian, and Toscana viruses. J Virol 81:5246–5256 [View Article][PubMed]
    [Google Scholar]
  3. Barr J. N., Rodgers J. W., Wertz G. W. 2006; Identification of the Bunyamwera bunyavirus transcription termination signal. J Gen Virol 87:189–198 [View Article][PubMed]
    [Google Scholar]
  4. Blakqori G., van Knippenberg I., Elliott R. M. 2009; Bunyamwera orthobunyavirus S-segment untranslated regions mediate poly(A) tail-independent translation. J Virol 83:3637–3646 [View Article][PubMed]
    [Google Scholar]
  5. Bouloy M., Hannoun C. 1976; Studies on lumbo virus replication. I. RNA-dependent RNA polymerase associated with virions. Virology 69:258–264 [View Article][PubMed]
    [Google Scholar]
  6. Bouloy M., Vialat P., Girard M., Pardigon N. 1984; A transcript from the S segment of the Germiston bunyavirus is uncapped and codes for the nucleoprotein and a nonstructural protein. J Virol 49:717–723[PubMed]
    [Google Scholar]
  7. Bouloy M., Pardigon N., Vialat P., Gerbaud S., Girard M. 1990; Characterisation of the 5′ and 3′ ends of viral messenger RNAs isolated from BHK21 cells infected with Germiston virus (Bunyavirus). Virology 175:50–58 [View Article]
    [Google Scholar]
  8. Cunningham C., Szilágyi J. F. 1987; Viral RNAs synthesized in cells infected with Germiston Bunyavirus. Virology 157:431–439 [View Article][PubMed]
    [Google Scholar]
  9. Dunn E. F., Pritlove D. C., Elliott R. M. 1994; The S RNA genome segments of Batai, Cache Valley, Guaroa, Kairi, Lumbo, Main Drain and Northway bunyaviruses: sequence determination and analysis. J Gen Virol 75:597–608 [View Article][PubMed]
    [Google Scholar]
  10. Dunn E. F., Pritlove D. C., Jin H., Elliott R. M. 1995; Transcription of a recombinant bunyavirus RNA template by transiently expressed bunyavirus proteins. Virology 211:133–143 [View Article][PubMed]
    [Google Scholar]
  11. Elliott R. M. 1985; Identification of nonstructural proteins encoded by viruses of the Bunyamwera serogroup (family Bunyaviridae). Virology 143:119–126 [View Article][PubMed]
    [Google Scholar]
  12. Elliott R. M. 1989a; Nucleotide sequence analysis of the small (S) RNA segment of Bunyamwera virus, the prototype of the family Bunyaviridae . J Gen Virol 70:1281–1285 [View Article][PubMed]
    [Google Scholar]
  13. Elliott R. M. 1989b; Nucleotide sequence analysis of the large (L) genomic RNA segment of Bunyamwera virus, the prototype of the family Bunyaviridae . Virology 173:426–436 [View Article][PubMed]
    [Google Scholar]
  14. Eshita Y., Ericson B., Romanowski V., Bishop D. H. 1985; Analyses of the mRNA transcription processes of snowshoe hare bunyavirus S and M RNA species. J Virol 55:681–689[PubMed]
    [Google Scholar]
  15. Fazakerley J. K., Gonzalez-Scarano F., Strickler J., Dietzschold B., Karush F., Nathanson N. 1988; Organization of the middle RNA segment of snowshoe hare Bunyavirus. Virology 167:422–432[PubMed] [CrossRef]
    [Google Scholar]
  16. Flick R., Pettersson R. F. 2001; Reverse genetics system for Uukuniemi virus (Bunyaviridae): RNA polymerase I-catalyzed expression of chimeric viral RNAs. J Virol 75:1643–1655 [View Article][PubMed]
    [Google Scholar]
  17. Ikegami T., Won S., Peters C. J., Makino S. 2007; Characterization of Rift Valley fever virus transcriptional terminations. J Virol 81:8421–8438 [View Article][PubMed]
    [Google Scholar]
  18. Jin H., Elliott R. M. 1993; Characterization of Bunyamwera virus S RNA that is transcribed and replicated by the L protein expressed from recombinant vaccinia virus. J Virol 67:1396–1404[PubMed]
    [Google Scholar]
  19. Lara E., Billecocq A., Leger P., Bouloy M. 2011; Characterization of wild-type and alternate transcription termination signals in the Rift Valley fever virus genome. J Virol 85:12134–12145 [View Article][PubMed]
    [Google Scholar]
  20. Leonard V. H., Kohl A., Osborne J. C., McLees A., Elliott R. M. 2005; Homotypic interaction of Bunyamwera virus nucleocapsid protein. J Virol 79:13166–13172 [View Article][PubMed]
    [Google Scholar]
  21. Li X., Palese P. 1994; Characterization of the polyadenylation signal of influenza virus RNA. J Virol 68:1245–1249[PubMed]
    [Google Scholar]
  22. Lowen A. C., Elliott R. M. 2005; Mutational analyses of the nonconserved sequences in the Bunyamwera Orthobunyavirus S segment untranslated regions. J Virol 79:12861–12870 [View Article][PubMed]
    [Google Scholar]
  23. Lowen A. C., Noonan C., McLees A., Elliott R. M. 2004; Efficient bunyavirus rescue from cloned cDNA. Virology 330:493–500 [View Article][PubMed]
    [Google Scholar]
  24. Masek T., Vopalensky V., Suchomelova P., Pospisek M. 2005; Denaturing RNA electrophoresis in TAE agarose gels. Anal Biochem 336:46–50 [View Article][PubMed]
    [Google Scholar]
  25. Osborne J. C., Elliott R. M. 2000; RNA binding properties of bunyamwera virus nucleocapsid protein and selective binding to an element in the 5′ terminus of the negative-sense S segment. J Virol 74:9946–9952 [View Article][PubMed]
    [Google Scholar]
  26. Patterson J. L., Holloway B., Kolakofsky D. 1984; La Crosse virions contain a primer-stimulated RNA polymerase and a methylated cap-dependent endonuclease. J Virol 52:215–222[PubMed]
    [Google Scholar]
  27. Pattnaik A. K., Abraham G. 1983; Identification of four complementary RNA species in Akabane virus-infected cells. J Virol 47:452–462[PubMed]
    [Google Scholar]
  28. Walter C. T., Barr J. N. 2011; Recent advances in the molecular and cellular biology of bunyaviruses. J Gen Virol 92:2467–2484 [View Article][PubMed]
    [Google Scholar]
  29. Weber F., Dunn E. F., Bridgen A., Elliott R. M. 2001; The Bunyamwera virus nonstructural protein NSs inhibits viral RNA synthesis in a minireplicon system. Virology 281:67–74 [View Article][PubMed]
    [Google Scholar]
  30. Whelan S. P., Barr J. N., Wertz G. W. 2004; Transcription and replication of nonsegmented negative-strand RNA viruses. Curr Top Microbiol Immunol 283:61–119[PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.042390-0
Loading
/content/journal/jgv/10.1099/vir.0.042390-0
Loading

Data & Media loading...

Most cited Most Cited RSS feed