1887

Journal of General Virology header logo

Volume 31, Issue 3

Defective-Interfering Particles of Semliki Forest Virus: Structural Differences between Standard Virus and Defective-Interfering Particles Free

Abstract

SUMMARY

Serial passaging of Semliki Forest virus in BHK cells at a constant input multiplicity of 50 p.f.u./cell resulted in a 4 log drop in the yield of infectious virus by passage 9. An interference analysis showed that this drop was due to the presence of defective-interfering (DI) particles. Attempts were made to separate the DI particles from standard virus by equilibrium and velocity centrifugation. Only equilibrium centrifugation on CsCl resolved the DI particles (identified by interference analyses) from standard virus. The buoyant density of the DI particles (1.23 g/ml) was higher than that of standard virus (ρ = 1·20 g/ml).

No difference was observed between the structural proteins of standard virus and DI particles. Analysis of the RNA of standard virus and DI particles showed that whereas standard virus contained only 42S RNA (mol. wt. approx. 4·2 × 10), DI particles contained two smaller pieces of RNA of mol. wt. 0.81 and 0·75 × 10 respectively. Infectivity assays showed that these low mol. wt. species were not only non-infectious but also interfered with the infectivity of 42S RNA from standard virus. Nucleocapsids derived from purified DI particles had a buoyant density 0·02 g/ml greater than the nucleocapsids from standard virus. Analysis of the RNA from DI nucleocapsids showed it to be entirely of the low mol. wt. class.

To account therefore for the density difference not only between DI particles and standard virus but also between their respective nucleocapsids we propose that each SFV DI particle contains several molecules of the low mol. wt. RNA species.

  • Received:
  • Accepted:
  • Published Online:
© Society for General Microbiology 1976
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-31-3-383
1976-06-01
2024-04-18
Loading full text...

Full text loading...

/deliver/fulltext/jgv/31/3/JV0310030383.html?itemId=/content/journal/jgv/10.1099/0022-1317-31-3-383&mimeType=html&fmt=ahah

References

  1. ACHESON N. M., TAMM I. 1970; Purification and properties of Semliki Forest virus nucleccapsids. Virology 41:306–320
     [Google Scholar]
  2. ATKINS G. J., SAMUELS J., KENNEDY S. I. T. 1974; Isolation and characterization of temperature-sensitive mutants of Sindbis virus strain AR339. Journal of General Virology 25:371–380
     [Google Scholar]
  3. CLEGG J. C. S., KENNEDY S. I. T. 1974a; Polyadenylic acid sequences in the virus RNA species of cells infected with Semliki Forest virus. Journal of General Virology 22:231–345
     [Google Scholar]
  4. CLEGG J. C. S., KENNEDY S. I. T. 1974b; In vitro synthesis of structural proteins of Semliki Forest virus directed by isolated 26 S RNA from infected cells. FEBS Letters 42:327–330
     [Google Scholar]
  5. CLEGG J. C. S., KENNEDY S. I. T. 1975; Translation of Semliki Forest virus intracellular 26S RNA: characterization of the products synthesised in vitro. European Journal of Biochemistry 53:175–183
     [Google Scholar]
  6. COMPANS R. W. 1971; Location of the glycoprotein in the membrane of Sindbis virus. Nature New Biology 229:114–116
     [Google Scholar]
  7. CRICK J., LEBEDEV A. I., STEWART D. L., BROWN F. 1966; The assay, extraction and storage of infective ribonucleic acid from foot-and-mouth disease virus. Journal of General Microbiology 43:59–70
     [Google Scholar]
  8. FRIEDMAN R. M., LEVY H. B., CARTER W. B. 1966; Replication of Semliki Forest virus: three forms of viral RNA produced during infection. Proceedings of the National Academy of Sciences of the United States of America 56:440–446
     [Google Scholar]
  9. GAROFF H., SIMONS K. 1974; Isolation and characterization of the membrane proteins of Semliki Forest virus. Virology 61:493–504
     [Google Scholar]
  10. HUANG A. S. 1973; Defective interfering viruses. Annual Review of Microbiology 27:101–119
     [Google Scholar]
  11. KENNEDY S. I. T. 1974; The effect of enzymes on structural and biological properties of Semliki Forest virus. Journal of General Virology 23:129–143
     [Google Scholar]
  12. KENNEDY S. I. T., BURKE D. C. 1972; Studies on the structural proteins of Semliki Forest virus. Journal of General Virology 14:87–98
     [Google Scholar]
  13. LAINE R., SÖDERLUND H., RENKONEN O. 1973; Chemical composition of Semliki Forest virus. Intervirology 1:110–118
     [Google Scholar]
  14. LEVIN J. G., FRIEDMAN R. M. 1971; Analysis of arbovirus ribonucleic acid forms by Polyacrylamide gel electrophoresis. Journal of Virology 7:504–514
     [Google Scholar]
  15. MARTIN B. A. B., BURKE D. C. 1974; Replication of Semliki Forest virus. I. Separation and characterization of the virus-specified RNA species formed in infected cells. Journal of General Virology 24:45–66
     [Google Scholar]
  16. MORSER M. J., KENNEDY S. I. T., BURKE D. C. 1973; Virus-specified polypeptides in cells infected with Semliki Forest virus. Journal of General Virology 21:19–29
     [Google Scholar]
  17. SCHLESINGER S., SCHLESINGER M., BÜRGE B. W. 1973; Defective virus particles from Sindbis virus. Virology 48:615–617
     [Google Scholar]
  18. SHENK T. E., STOLLAR V. 1973; Defective-interfering particles of Sindbis virus. Virology 53:162–173
     [Google Scholar]
  19. SIMMONS D. T., STRAUSS J. H. 1972; Replication of Sindbis virus. I. Relative size and genetic content of 26S and 49S RNA. Journal of Molecular Biology 71:599–613
     [Google Scholar]
  20. WALTERS S., BURKE D. C., SKEHEL J. J. 1967; Interferon production and RNA inhibitors. Journal of General Virology 1:349–362
     [Google Scholar]
  21. WEISS B., SCHLESINGER S. 1973; Defective interfering passages of Sindbis virus: chemical composition, biological activity and mode of interference. Journal of Virology 12:862–871
     [Google Scholar]
  22. WENGLER G., BEATO M., HACKEMACK B. A. 1974; Translation of 26S virus-specific RNA from Semliki Forest virus-infected cells in vitro. Virology 61:120–128
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-31-3-383
Loading
Defective-Interfering Particles of Semliki Forest Virus: Structural Differences between Standard Virus and Defective-Interfering Particles
J. Gen. Virol. 31, 383 (1976); https://doi.org/10.1099/0022-1317-31-3-383
/content/journal/jgv/10.1099/0022-1317-31-3-383
/content/journal/jgv/10.1099/0022-1317-31-3-383
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