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Generation of a Defective RNA of Avian Coronavirus Infectious Bronchitis Virus (IBV)

Defective RNA of Coronavirus IBV
  • Zoltan Penzes
  • Kefford W. Tibbles
  • Kathy Shaw
  • Paul Britton
  • T. David K. Brown
  • David Cavanagh
Chapter
  • 209 Downloads
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 380)

Abstract

The Beaudette strain of IBV was passaged 16 times in chick kidney (CK) cells. Total cellular RNA was analyzed by Northern hybridization and was probed with 32P-labeled cDNA probes corresponding to the first 2 kb of the 5′ end of the genome, but excluding the leader, and to the last 1.8 kb of the 3′ end of the genome. A new, defective IBV RNA species (CD-91) was detected at passage six. The defective RNA, present in total cell extract RNA and in oligo-(dT)30-selected RNA from passage 15, was amplified by the reverse transcription-polymerase chain reaction (RT-PCR) to give four fragments. The oligonucleotides used were selected such that CD-91 RNA, but not the genomic RNA, would be amplified. Cloning and sequencing of the PCR products showed that CD-91 comprises 9.1 kb and has three regions of the genome. It contains 1133 nucleotides from the 5′ end of the genome, 6322 from gene lb corresponding to position 12423 to 18744 in the IBV genome and 1626 from the 3’ end of the genome. At position 749 one nucleotide, an adenine residue, was absent from CD-91 RNA. By Northern hybridization CD-91 RNA was detected in virions in higher amounts than the subgenomic mRNAs.

Keywords

Vero Cell Infectious Bronchitis Virus Northern Hybridization Adenine Residue Guanidinium Isothiocyanate 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Baric, R. S., Stohlman, S. A., Razavi, M. K., and Lai, M. M. C. (1985). Characterisation of leader related small RNAs in coronavirus infected cells. Further evidence for leader primed mechanism transcription. Virus Res. 3, 19–33.Google Scholar
  2. 2.
    Baric, R. S., Shieh, C., Stohlman, S. A., and Lai, M. M. C. (1987). Analysis of intracellular small RNAs of mouse hepatitis virus: evidence for discontinuous transcription. Virology 156, 342–354.PubMedCrossRefGoogle Scholar
  3. 3.
    Boursnell, M. E. G., Brown, T. D. K., Foulds, I. J., Green, P. F., Tomley, F. M., and Binns, M. M. (1987). Completion of the sequence of the genome of the coronavirus avian infectious bronchitis virus. J. Gen. Virol. 68, 57–77.PubMedCrossRefGoogle Scholar
  4. 4.
    Cavanagh, D., Davis, P. J., and Cook, J. K. A. (1992). Infectious bronchitis virus: evidence for recombination within the Massachusetts serotype. Avian Pathol. 21,401–408.PubMedCrossRefGoogle Scholar
  5. 5.
    de Groot, R. J., van der Most, R. G., and Spaan, W. J. M. (1992). The fitness of defective interfering murine coronavirus DI-a and its derivatives is decreased by nonsense and frameshift mutations. J. Virol.66, 5898–5905.PubMedGoogle Scholar
  6. 6.
    Fosmire, J. A., Hwang, K., and Makino, S. (1992). Identification and characterization of a coronavirus packaging signal. J. Virol. 66, 3522–3530.PubMedGoogle Scholar
  7. 7.
    Furuya, T., Macnaughton, T. B., La Monica, N., and Lai, M. M. C. (1993). Natural evolution of coronavirus defective-interfering RNA involves RNA recombination. Virology 194, 408–413.PubMedCrossRefGoogle Scholar
  8. 8.
    Jeong, Y. S., and Makino, S. (1994). Evidence for coronavirus discontinuous transcription. J. Virol. 68, 2615–2623.PubMedGoogle Scholar
  9. 9.
    Kim, Y., Jeong, Y., and Makino, S. (1993). Analysis of cis-acting sequences essential for coronavirus defective interfering RNA replication. Virology 197, 53–63.PubMedCrossRefGoogle Scholar
  10. 10.
    Lai, M. M. C., Baric, R. S., Makino, S., Keck, J. G., Egbert, J., Leibowitz, J. L., and Stohlman, S. A. (1985). Recombination between nonsegmented RNA genomes of murine coronaviruses. J. Virol. 56,449–456.PubMedGoogle Scholar
  11. 11.
    Lin, Y., and Lai, M. M. C. (1993). Deletion mapping of a mouse hepatitis virus defective interfering RNA reveals the requirement of an internal and discontiguous sequence for replication. J. Virol. 67,6110–6118.PubMedGoogle Scholar
  12. 12.
    Makino, S., Taguchi, F., and Fujiwara, K. (1984). Defective interfering particles of mouse hepatitis virus.Virology 133, 9–17.PubMedCrossRefGoogle Scholar
  13. 13.
    Makino, S., Fujioka, N., and Fujiwara, K. (1985). Structure of the intracellular viral RNAs of defective interfering particles of mouse hepatitis virus. J. Virol 54, 329–336.PubMedGoogle Scholar
  14. 14.
    Makino, S., Shieh, C., Soe, L., Baker, S. C, and Lai, M. M. C. (1988). Primary structure and translation of a defective interfering RNA of murine coronavirus. Virology 166, 550–560.PubMedCrossRefGoogle Scholar
  15. 15.
    Makino, S., Yokomori, K., and Lai, M. M. C. (1990). Analysis of efficiently packaged defective interfering RNAs of murine coronavirus: localization of a possible RNA-packaging signal. J. Virol 64, 6045–6053.PubMedGoogle Scholar
  16. 16.
    Sethna, P. B., Hofmann, M. A., and Brian, D. A. (1991). Minus-strand copies of replicating coronavirus mRNAs contain antileaders. J. Virol 65, 320–325.PubMedGoogle Scholar
  17. 17.
    van der Most, R. G., Bredenbeek, P. J., and Spaan, W. J. M. (1991). A domain at the 3’ end of the polymerase gene is essential for encapsidation of coronavirus defective interfering RNAs. J. Virol 65,3219–3226.PubMedGoogle Scholar
  18. 18.
    Zhao, X., Shaw, K., and Cavanagh, D. (1993). Presence of subgenomic mRNAs in virions of coronavirus IBV. Virology 196, 172–178.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • Zoltan Penzes
    • 1
  • Kefford W. Tibbles
    • 2
  • Kathy Shaw
    • 1
  • Paul Britton
    • 1
  • T. David K. Brown
    • 2
  • David Cavanagh
    • 1
  1. 1.Division of Molecular BiologyInstitute for Animal HealthCompton, NewburyUK
  2. 2.Division of Virology, Department of PathologyUniversity of CambridgeCambridgeUK

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