1887

Abstract

Defective interfering (DI) RNA molecules derived from the genomic L RNA segment of tomato spotted wilt virus (TSWV) were generated during sequential passage of the virus at high multiplicity. Characterization of DI RNAs from four distinct isolates by Northern blot analysis and sequence determination revealed that both the 5′ and 3′ genomic termini were retained in these molecules. Each DI RNA contained a single internal deletion of approximately 60% to 80% of the L RNA segment. All DI RNAs studied maintain an open reading frame (ORF) which suggests that these defective molecules should be translatable by ribosomes. Detection of only defective molecules with ORFs indicates either that association with ribosomes or translation is a prerequisite for the selection and maintenance of replicating DI RNAs, or that the truncated proteins produced play a role in their selection or replication. Analysis of the junction sites in the DI RNAs showed that short nucleotide sequences are repeated, one at the release and another at the reinitiation point on the L RNA. One of these is lost during the generation of the DI molecules. The presence of repeated sequences at the junction sites seems to be unique for tospovirus DI L RNAs; they have not been described for other DI systems of either positive- or negative-strand RNA viruses. A model for TSWV DI RNA generation is proposed in which the viral polymerase can ‘jump’ across the internal sequences from one secondary structure to another containing the repeated sequences, during the replication of the viral complementary L RNA segment.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-73-10-2509
1992-10-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/jgv/73/10/JV0730102509.html?itemId=/content/journal/jgv/10.1099/0022-1317-73-10-2509&mimeType=html&fmt=ahah

References

  1. Amesse L. S., Pridgen C. L., Kingsbury D. W. 1982; Sendai virus DI RNA species with conserved virus genome termini and extensive internal deletions. Virology 118:17–27
    [Google Scholar]
  2. Bailey J. M., Davidson N. 1976; Methylmercury as a reversible denaturating agent for agarose gel electrophoresis. Analytical Biochemistry 70:75–85
    [Google Scholar]
  3. Baric R. S., Shieh C. K., Stohlman S. A., Lai M. M. C. 1987; Analysis of intracellular small RNAs of mouse hepatitis virus: evidence for discontinuous transcription. Virology 156:342–354
    [Google Scholar]
  4. Bujarski J. J., Dzianot A. M. 1991; Generation and analysis of nonhomologous RNA-RNA recombinants of brome mosaic virus: sequence complementarities at crossover sites. Journal of Virology 65:4153–4159
    [Google Scholar]
  5. Burgyan J., Rubino L., Russo M. 1991; De now generation of cymbidium ringspot virus defective interfering RNA. Journal of General Virology 72:505–509
    [Google Scholar]
  6. Cascone P. J., Carpenter C. D., Li X. H., Simon A. E. 1990; Recombination between satellite RNAs of turnip crinkle virus. EMBO Journal 9:1709–1715
    [Google Scholar]
  7. Cunningham C., Szilagyi J. F. 1987; Viral RNAs synthesized in cells infected with Germiston bunyavirus. Virology 157:431–439
    [Google Scholar]
  8. de Ávila A. C., Huguenot C., Resende R., de O., Kitajima E. W., Goldbach R., Peters D. 1990; Serological differentiation of 20 isolates of tomato spotted wilt virus. Journal of General Virology 71:2801–2807
    [Google Scholar]
  9. de Ávila A. C., de Haan P., Smeets M. L. L., Resende R. de O., Kormelink R., Kitajima E. W., Goldbach R., Peters D. 1992; Distinct levels of relationships between tospovirus isolates. Archives of Virology (in press)
    [Google Scholar]
  10. de Haan P., Wagemakers L., Peters D., Goldbach R. 1990; The S RNA segment of tomato spotted wilt virus has an ambisense character. Journal of General Virology 71:1001–1007
    [Google Scholar]
  11. de Haan P., Kormelink R., Resende R., de O., van Poelwijk F., Peters D., Goldbach R. 1991; Tomato spotted wilt virus L RNA encodes a putative RNA polymerase. Journal of General Virology 72:2207–2216
    [Google Scholar]
  12. Holland J. J. 1985; Generation and replication of defective viral genomes. In Virology pp. 77–99 Edited by Fields B. N., Knipe D. M. New York: Raven Press;
    [Google Scholar]
  13. Jennings P. A., Finch J. T., Winter G., Robertson J. S. 1983; Does the higher order structure of the influenza virus ribonucleo-protein guide sequence rearrangements in the influenza viral RNA?. Cell 34:619–627
    [Google Scholar]
  14. Kirkegaard K., Baltimore D. 1986; The mechanism of RNA recombination in poliovirus. Cell 47:433–443
    [Google Scholar]
  15. Knorr D. A., Mullin R. H., Hearne P. Q., Morris A. T. 1991; De now generation of defective interfering RNAs of tomato bushy stunt virus by high multiplicity passage. Virology 181:193–202
    [Google Scholar]
  16. Kozak M. 1978; How do eucaryotic ribosomes select initiation regions in messenger RNA?. Cell 15:1109–1123
    [Google Scholar]
  17. Kozak M. 1989; The scanning model for translation: an update. Journal of Cell Biology 108:229–241
    [Google Scholar]
  18. Kuge S., Saito I., Nomoto A. 1986; Primary structures of poliovirus defective interfering particles genomes and possible generation mechanisms of the particles. Journal of Virology 192:473–487
    [Google Scholar]
  19. Lazzarini R. A., Keene J. D., Schubert M. 1981; The origins of defective interfering particles of the negative-strand RNA viruses. Cell 26:145–154
    [Google Scholar]
  20. Maiss E., Ivanova L., Breyel E., Adam G. 1991; Cloning and sequencing of the S RNA from a Bulgarian isolate of tomato spotted wilt virus. Journal of General Virology 72:461–464
    [Google Scholar]
  21. Makino S., Shieh C. K., Sob L. H., Baker S. C., Lai M. M. C. 1988; Primary structure and translation of a defective interfering RNA of murine coronavirus. Virology 166:550–560
    [Google Scholar]
  22. Maniatis T., Fritsch R. F., Sambrook J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  23. Marchuk D., Drumm M., Saulino A., Collins F. S. 1991; Construction of T-vectors, a rapid and general system for direct cloning of unmodified PCR products. Nucleic Acids Research 19:1154
    [Google Scholar]
  24. Meier E., Harmison G. G., Keene J. D., Schubert M. 1984; Sites of copy choice replication involved in the generation of vesicular stomatitis virus defective-interfering particle RNAs. Journal of Virology 51:515–521
    [Google Scholar]
  25. Morris T. J., Knorr D. A. 1990; Defective interfering viruses associated with plant virus infection. In New Aspects of Positive-Strand RNA Viruses pp. 123–127 Edited by Brinton M. A., Heinz F. X. Washington, D.C.: American Society for Microbiology;
    [Google Scholar]
  26. Nayak D. P., Chambers T. M., Akkina R. K. 1985; Defective-interfering (DI) RNAs of influenza viruses: origin, structure, expression and interfering. Current Topics in Microbiology and Immunology 114:103–151
    [Google Scholar]
  27. Nayak D. P., Chambers T. M., Akkina R. K. 1990; Structure of defective interfering RNAs of influenza viruses and their role in interference. In The Influenza Viruses pp. 269–317 Edited by Krug R. M. New York: Plenum Press;
    [Google Scholar]
  28. Patel A. H., Elliott R. M. 1992; Characterization of Bunyamwera virus defective interfering particles. Journal of General Virology 73:389–396
    [Google Scholar]
  29. Perrault J. 1981; Origin and replication of defective interfering particles. Current Topics in Microbiology and Immunology 93:151–207
    [Google Scholar]
  30. Re G. G., Morgan E. M., Kingsbury D. W. 1985; Nucleotide sequences responsible for generation of internally deleted Sendai virus defective interfering genomes. Virology 146:27–37
    [Google Scholar]
  31. Resende R. De O., de Haan P., de Ávila A. C., Kitajima E. W., Kormelink R., Goldbach R., Peters D. 1991; Generation of envelope and defective interfering RNA mutants of tomato spotted wilt virus by mechanical passage. Journal of General Virology 72:2375–2383
    [Google Scholar]
  32. Romanova L. I., Blinov V. M., Tolskaya E. A., Vktorova E. G., Kolesnikowa M. S., Guseva E. A., Agol V. I. 1986; The primary structure of crossover regions of intertypic poliovirus recombinants: a model of recombination between RNA genomes. Virology 155:202–213
    [Google Scholar]
  33. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences, U.S.A. 74:5463–5467
    [Google Scholar]
  34. Scallan M. F., Elliott R. M. 1992; Defective RNAs in mosquito cells persistently infected with Bunyamwera virus. Journal of General Virology 73:53–60
    [Google Scholar]
  35. Sivasubramanian N., Nayak D. P. 1983; Defective interfering influenza RNAs of polymerase 3 gene contain single as well as multiple internal deletions. Virology 124:232–237
    [Google Scholar]
  36. Yang F., Lazzarini R. A. 1983; Analysis of the recombination event generating a vesicular stomatitis virus deletion defective interfering particle. Journal of Virology 45:766–772
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-73-10-2509
Loading
/content/journal/jgv/10.1099/0022-1317-73-10-2509
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error