1887

Abstract

The cDNA sequence of genomic segment 5 of bovine rotavirus (RF strain) has been inserted into baculovirus transfer vectors, downstream of the polyhedrin promoter. Recombinant baculoviruses containing gene 5 were selected and the protein was expressed to high yields in cells. The recombinant protein was inoculated into rabbits and mice to produce specific hyperimmune antisera. The polyclonal antiserum reacted with a protein in rotavirus-infected MA104 cells and with a protein translated . This serum was also used to confirm that the gene 5 protein is not a structural protein. Recently, the gene 5 product has been predicted to be a zinc finger protein and reported to contain a highly conserved arrangement of cysteine residues; here, we demonstrate that the recombinant gene 5 protein binds zinc and is an RNA-binding protein as are several other zinc finger proteins.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-73-8-1931
1992-08-01
2022-08-13
Loading full text...

Full text loading...

/deliver/fulltext/jgv/73/8/JV0730081931.html?itemId=/content/journal/jgv/10.1099/0022-1317-73-8-1931&mimeType=html&fmt=ahah

References

  1. Bican P., Cohen J., Charpilienne A., Scherrer R. 1982; Purification and characterization of bovine rotavirus cores. Journal of Virology 43:1113–1117
    [Google Scholar]
  2. Both G. W., Bellamy A. R., Street J. E., Siegman L. J. 1982; A general strategy for cloning double-stranded RNA: nucleotide sequence of the simian 11 rotavirus gene 8. Nucleic Acids Research 10:7075–7088
    [Google Scholar]
  3. Boyle J. F., Holmes K. V. 1986; RNA-binding proteins of bovine rotavirus. Journal of Virology 58:561–568
    [Google Scholar]
  4. Brémont M., Charpilienne A., Chabanne D., Cohen J. 1987; Nucleotide sequence and expression in Escherichia coli of the gene encoding the non-structural protein NCVP2 of bovine rotavirus. Viology 161:138–144
    [Google Scholar]
  5. Brémont M., Vautherot D., Charpilienne A., Brother P., Cohen J. 1990; Genome comparison of group A and group C rotaviruses. Third International Symposium on Double-stranded Viruses, Hawaii
    [Google Scholar]
  6. Cohen J. 1977; Ribonucleic acid polymerase activity associated with purified calf rotavirus. Journal of General Virology 36:395–402
    [Google Scholar]
  7. Cohen J., Charpilienne A., Chilmonczyk S., Estes M. K. 1989; Nucleotide sequence of bovine rotavirus gene 1 and expression of the gene product in baculovirus. Virology 171:131–140
    [Google Scholar]
  8. Estes M. K., Cohen J. 1989; Rotavirus gene structure and function. Microbiological Reviews 53:410–449
    [Google Scholar]
  9. Evans R. M., Hollenberg S. M. 1988; Zinc fingers: gilt by association. Cell 52:1–3
    [Google Scholar]
  10. Frankel A. D., Bredt D. S., Pabo C. O. 1988; Tat protein from human immunodeficiency virus forms a metal-linked dimer. Sciences 240:70–73
    [Google Scholar]
  11. Gallegos C. O., Patton J. T. 1989; Characterization of rotavirus replication intermediates. A model for the assembly of single-shelled particles. Virology 172:616–627
    [Google Scholar]
  12. Huismans H., Joklik W. K. 1976; Reovirus-coded polypeptides in infected cells: isolation of two native monomeric polypeptides with affinity for single-stranded and double-stranded RNA, respectively. Virology 70:411–424
    [Google Scholar]
  13. Huismans H., Van Dikj W. K., Bauskin A. R. 1987; In vitro phosphorylation and purification of a non-structural protein of bluetongue virus with affinity for single-strand RNA. Journal of Virology 61:3589–3595
    [Google Scholar]
  14. Johnson M. A., McCrae M. A. 1989; Quantitative analysis of regulation of gene expression during virus replication. Journal of Virology 63:2048–2055
    [Google Scholar]
  15. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  16. L’Haridon R., Scherrer R. 1976; In vitro culture of a rotavirus associated with neonatal calf scours. Annales de Recherches Vétérin-aires 7:373–381
    [Google Scholar]
  17. Loeber G., Parson R., Tegtmeyer P. 1989; The zinc finger region of simian virus 40 large T antigen. Journal of Virology 63:94–100
    [Google Scholar]
  18. Luckow V. A., Summers M. D. 1988; High level expression of non-fused foreign genes with Autographa califomica nuclear polyhedrosis virus expression vectors. Virology 170:31–39
    [Google Scholar]
  19. McCrae M. A., McCorquodale J. G. 1982; The molecular biology of rotaviruses. II. Identification of the protein-coding assignments of calf rotavirus genome RNA species. Virology 117:435–443
    [Google Scholar]
  20. Meric C., Goff S. P. 1989; Characterization of Moloney murine leukemia virus mutants with single amino acid substitutions in the Cys-His box of the nucleocapsid protein. Journal of Virology 63:1558–1568
    [Google Scholar]
  21. Miller J., McLachlan A. D., Klug A. 1985; Repetitive zinc-binding domains in the protein transcription factor IIIA from Xenopus oocytes. EMBO Journal 4:1609–1614
    [Google Scholar]
  22. Mitchell D. B., Both G. W. 1990; Conservation of a potential metal binding motif despite extensive sequence diversity in the rotavirus nonstructural protein NS53. Virology 174:618–621
    [Google Scholar]
  23. Roy P. 1989; Bluetongue virus genetics and genome structure. Virus Research 13:179–206
    [Google Scholar]
  24. Schiff L. A., Nibert M. L., Man Sung Co , Brown E. G., Fields B. N. 1988; Distinct binding sites for zinc and double-stranded RNA in the reovirus outer capsid protein σ 3. Molecular and Cellular Biology 8:273–283
    [Google Scholar]
  25. Summers M. D., Smith G. E. 1987; A manual of methods for baculovirus vectors and insect cell culture procedures. Texas Agricultural Experiment Station Bulletin no. 1555
    [Google Scholar]
  26. Thomas C. P., Booth T. F., Roy P. 1990; Synthesis of bluetongue virus-encoded phosphoprotein and formation of inclusion bodies by recombinant baculovirus in insect cells: it binds the single-stranded RNA species. Journal of General Virology 71:2073–2083
    [Google Scholar]
  27. Towbin H., Staehelin T., Gordon I. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, U.S.A. 76:4350–4354
    [Google Scholar]
  28. Waitukaitis J. L., Robbins J. B., Nieschlag E. 1971; A method for producing specific antisera with small doses of immunogen. Production of antisera with small doses of immunogen. Journal of Clinical Endocrinology 33:988–991
    [Google Scholar]
  29. Wiener J. R., Joklik W. K. 1987; Comparison of the reovirus serotype 1, 2, and 3 S3 genome segments encoding the nonstructural protein σ NS. Virology 161:332–339
    [Google Scholar]
  30. Wiener J. R., Bartlett J. A., Joklik W. K. 1989; The sequences of reovirus serotype 3 genome segments M1 and M3 encoding the minor protein mu 2 and the major nonstructural protein mu NS, respectively. Virology 169:293–304
    [Google Scholar]
  31. Xu L., Harbour D., McCrae M. A. 1990; Analysis of sequence diversity in gene 5 of rotaviruses. Third International Symposium on Double-stranded Vineses Hawaii:
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-73-8-1931
Loading
/content/journal/jgv/10.1099/0022-1317-73-8-1931
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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