1887

Journal of General Virology header logo

Volume 43, Issue 1

Studies of the Replication of a Bovine Enterovirus RNA Free

Abstract

SUMMARY

Replicative intermediate (RI), replicative form (RF) and single-stranded (SS) RNA have been isolated from BHK cells infected with a bovine enterovirus by salt precipitation and gel filtration techniques. Kinetic experiments showed that at no time up to 16 h post-infection (p.i.) did the amount of RF exceed that of RI or SS RNA.

Electrophoresis of RF on 1.5% polyacrylamide-agarose gels showed that at least three species of double-stranded RNA were present, one of which was associated with an accessible poly(A)-containing tract. All of the RF was denatured by 99% dimethylsulphoxide (DMSO), although reannealling occurred rapidly when samples were returned to aqueous conditions. No evidence for circular structures in the RF molecular population was found by use of caesium sulphate density gradients containing ethidium bromide.

Treatment of RI with ribonuclease produced double-stranded RNA molecules, some of which were smaller in size than intact RF. Denaturation with DMSO and analysis on 99% DMSO sucrose gradients showed that the RI did not contain single strands of greater length than virion RNA. A portion of the RI bound to poly(U)-Sepharose 4B columns. The poly(A) tracts involved were present only in the nascent RNA strands with greatest sedimentation coefficients (30 to 35S).

Bovine enterovirus induced SS RNA was heterogeneous with regard to both sedimentation through sucrose gradients and mobility on acrylamide gels compared to purified virion RNA. The reason for this difference has never been satisfactorily resolved. Sedimentation through 99% DMSO-sucrose gradients showed that the heterogeneity was due to aggregation rather than any variation in chain length or conformational differences.

Our results support the single-stranded template model rather than a circular model for picornavirus RNA replication.

  • Received:
  • Accepted:
  • Published Online:
© Journal of General Virology 1979
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-43-1-75
1979-04-01
2024-04-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/43/1/JV0430010075.html?itemId=/content/journal/jgv/10.1099/0022-1317-43-1-75&mimeType=html&fmt=ahah

References

  1. Agol V. I., Romanova L. I., Cumakov I. M., Dunaevskaya L. D., Bogdanov A. A. 1972; Circularity and cross-linking in preparations of replicative form of encaphalomyocarditis virus RNA. Journal of Molecular Biology 72:77–89
     [Google Scholar]
  2. Agol V. I., Romanova L. I., Tolskaya E. A., Bogdanov A. A. 1969; Some properties of the replicative form of the RNA of mouse encaphalomyocarditis virus. Dokladii Akademii 189:428–431
     [Google Scholar]
  3. Baltimore D. 1969; The replication of picornaviruses. In The Biochemistry of Viruses pp 129–160 Edited by Levy H. B. New York: Marcel Dekker;
     [Google Scholar]
  4. Bishop J. M., Levintow L. 1971; Replicative forms of viral RNA. Progress in Medical Virology 13:1–82
     [Google Scholar]
  5. Brown F., Martin S. J. 1965; A new model for virus ribonucleic acid replication. Nature, London 208:861–863
     [Google Scholar]
  6. Clements J. B., Martin S. J. 1971; Evidence for large strands of ribonucleic acid induced by a bovine enterovirus. Journal of General Virology 12:221–232
     [Google Scholar]
  7. Goldstein N. O., Pardoe I. U., Burness A. T. H. 1976; Requirement of an adenylic-acid rich segment for the infectivity of encapholomyocarditis virus RNA. Journal of General Virology 31:271–276
     [Google Scholar]
  8. Koliais S. L., Dimmock N. J. 1973; Replication of rhinovirus RNA. Journal of General Virology 20:1–15
     [Google Scholar]
  9. Lundquist R. E., Maizel J. V. Jun 1978; Structural studies on the RNA component of the poliovirus replication complex. Virology 85:434–444
     [Google Scholar]
  10. McFerran J. B. 1962; Bovine enteroviruses. Annals of the New York Academy of Sciences 101:436–443
     [Google Scholar]
  11. Macpherson L. A., Stoker M. G. P. 1962; Polyoma transformation of hamster cell clones –an investigation of genetic factors affecting cell competence. Virology 16:147–151
     [Google Scholar]
  12. Martin S. J., Johnston M. D., Clements J. B. 1970; Purification and characterization of bovine enteroviruses. Journal of General Virology 7:103–113
     [Google Scholar]
  13. Martin S. J., Ter Meulen V. 1976; A rapid method for the quantitative study of RNA from canine distemper virus infected cells. Journal of General Virology 32:321–325
     [Google Scholar]
  14. Meyer J., Lundquist R. E., Maizel J. V. Jun 1978; Structural studies on the RNA component of the poliovirus replication complex. Virology 85:445–455
     [Google Scholar]
  15. Miller R. L., Plageman P. G. W. 1972; Purification of mengovirus and identification of an A-rich segment in its ribonucleic acid. Journal of General Virology 17:349–353
     [Google Scholar]
  16. Montagnier L., Sanders F. K. 1963; Replicative form of encephalomyocarditis virus ribonucleic acid. Nature, London 199:664–667
     [Google Scholar]
  17. Noble J., Levintow L. 1970; Dynamics of poliovirus-specific RNA synthesis and the effects of inhibitors of virus replication. Virology 40:634–642
     [Google Scholar]
  18. Planterose D. N., Birch P. J., Pilch D. J. F., Sharpe T. J. 1970; Antiviral activity of double-stranded RNA and virus-like particles from Penicillium stoloniferum. Nature, London 227:504–505
     [Google Scholar]
  19. Szybalski W. 1968; Use of caesium sulphate for equilibrium density gradient centrifugation. In Methods in Enzymology vol 12B pp 330–360 Edited by Grossman L., Moldave K. New York and London: Academic Press;
     [Google Scholar]
  20. Thach S. S., Dobbertin D., Lawrence C., Golini F., Thach R. E. 1974; The mechanism of viral replication. Structure of replication complexes of encephalomyocarditis virus. Proceedings of the National Academy of Sciences of the United States of America 71:2549–2553
     [Google Scholar]
  21. Todd D., Martin S. J. 1975; Determination of the molecular weight of bovine enterovirus RNA by nuclease digestion. Journal of General Virology 26:121–129
     [Google Scholar]
  22. Wall R., Taylor M. W. 1970; Mengovirus RNA synthesis in productive and restrictive cell lines. Virology 42:78–86
     [Google Scholar]
  23. Wild T. F., Martin S. J., Brown F. 1968; A study of the heterogeneous 37S ribonucleic acid induced by foot-and-mouth disease virus. Biochemical Journal 107:395–401
     [Google Scholar]
  24. Yogo Y., Wimmer E. 1973; Poly(A) and poly(U) in poliovirus double stranded RNA. Nature, New Biology 242:171–174
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-43-1-75
Loading
Studies of the Replication of a Bovine Enterovirus RNA
J. Gen. Virol. 43, 75 (1979); https://doi.org/10.1099/0022-1317-43-1-75
/content/journal/jgv/10.1099/0022-1317-43-1-75
/content/journal/jgv/10.1099/0022-1317-43-1-75
Loading

Data & Media loading...

Most cited 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