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Interactions between the IBV Nucleocapsid Protein and RNA Sequences Specific for the 3′ end of the Genome

  • Ellen W. Collisson
  • Anna K. Williams
  • Shan-Ing Chung
  • Minglong Zhou
Chapter
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 380)

Abstract

The infectious bronchitis virus (IBV) nucleocapsid protein was expressed as a fusion protein in bacteria. The coding sequence differed from the native protein only in the addition of six histidine residues at the amino terminus which were used for enrichment with a nickel affinity column. In gel shift assays, the mobility of labelled G RNA was decreased with increasing concentrations of the fusion protein. Competitive gel shift assays with labelled G RNA indicated that the protein interacted with relatively high avidities to several unlabelled RNAs representing sequences at the 3′ noncoding end of the IBV genome. Cache Valley virus (a bunyavirus) mRNA transcribed from the smaller segment cDNA also inhibited the interaction with IBV G RNA to the same extent as homologous unlabelled G RNA. In contrast, interactions of the fusion protein with a region from 99 to 249 bases from the 3’ terminus of the IBV genome and bovine liver RNA were relatively weak. The binding of IBV nucleocapsid protein with RNA probably requires specific sequences and/or structures that are present at a number of sites on the genome, and may represent a common mechanism used by similar viral proteins whose functions depend on binding to RNA.

Keywords

Fusion Protein Infectious Bronchitis Virus Nucleocapsid Protein High Avidity Mouse Hepatitis Virus 
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.
    Boursnell, M. E. G., Binns, M. M., Foulds, I. J., and Brown, T. D. K. (1985). Sequences of the nucleocapsid genes from two strains of avian infectious bronchitis virus. J. Gen. Virol. 66, 573–580.PubMedCrossRefGoogle Scholar
  2. 2.
    Williams, A. K., Wang, L, Sneed, L. W., and Collisson, E. W. (1992). Comparative analyses of the nucleocapsid genes of several strains of infectious bronchitis virus and other coronaviruses. Virus Res. 25,213–222.PubMedCrossRefGoogle Scholar
  3. 3.
    Masters, P. S. (1992). Localization of an RNA-binding domain in the nucleocapsid protein of the coronavirus mouse hepatitis virus. Archives of Virol ,125, 141–160.CrossRefGoogle Scholar
  4. 4.
    Nelson, G. W., and Stohlman, S. A. (1993). Localization of the RNA-binding domain of mouse hepatitis virus nucleocapsid protein. J. Gen. Virol. 74, 1975–1979.PubMedCrossRefGoogle Scholar
  5. 5.
    Stohlman, S. A., Baric, R. S., Nelson, G. N., Soe, L. H., Welter, L. M., and Deans, R. J. (1988). Specific interaction between coronavirus leader RNA and nucleocapsid protein. J. Virol. 62(11), 4288–4295.PubMedGoogle Scholar
  6. 6.
    Sambrook, S., Fritsch, E. F., and Maniatis, T. (1989). “Molecular cloning-a laboratory manual.” 2nd edition. Cold Spring Harbor Laboratory Press.Google Scholar
  7. 7.
    Sneed, L. W, Butcher, G. D., Parr, R., Wang, L., and Collisson, E. W. (1989). Comparisons of the structural proteins of avian infectious bronchitis virus as determined by western blot analysis. Viral Immunol. 2(3), 221–227.CrossRefGoogle Scholar
  8. 8.
    Parr, R. L., and Collisson, E. W. 1993. Epitopes on the spike protein of a nephropathogenic strain of infectious bronchitis virus. Arch Virol. 133, 369–383.PubMedCrossRefGoogle Scholar
  9. 9.
    Chung, S.-I. (1992). Molecular and Pathogenesis Studies of Cache Valley Virus: A Common Teratogenic Agent in Sheep. Dissertation Thesis. Department of Veterinary Microbiology, Texas A&M University, College Station, TX.Google Scholar
  10. 10.
    Chodosh, L. A. (1992). Chapter 12.2. DNA-Protein interactions: Mobility shift DNA-binding assay using gel electrophoresis. Current Protocols, Supplement 3. Editors. F.M. Ausubel, R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, S.A. Smith & K. Struhl. Wiley & Sons, New York.Google Scholar

Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • Ellen W. Collisson
    • 1
  • Anna K. Williams
    • 2
  • Shan-Ing Chung
    • 3
  • Minglong Zhou
    • 1
  1. 1.Department of Veterinary PathobiologyTexas A&M UniversityCollege StationUSA
  2. 2.The Scripps Research InstituteLa JollaUSA
  3. 3.Molecular Biology DivisionDevelopment Center for BiotechnologyTaipeiTaiwan

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