1887

Abstract

SUMMARY

Purified Sendai virus nucleocapsids isolated from infected cells were used to programme a transcription system to study virus-specific RNA synthesis. The RNA products were analysed for size by centrifugation before and after denaturation with formamide or glyoxal. The polarity of the products [message (+) or genome (-) strands] was analysed by RNA-RNA hybridization. The non-denatured RNA products sedimented in three groups: 7S to 22S single-stranded RNA transcripts and two partially ribonuclease-resistant complexes. One complex, representing 12% of the total product, sedimented at 26S to 36S. After denaturing the 26S to 36S complex to single-stranded molecules, about half of the RNAs sedimented at 25S to 54S and about half at 6S to 24S. The second complex, representing about 13% of the total RNA product, sedimented at 42S to 52S. After denaturing, about 10% of the single-stranded RNAs sedimented at 38S to 52S and about 90% sedimented at 6S to 19S. In hybridization studies, single-stranded RNAs that sedimented at < 19S were predominantly of message sense (+ strand), whereas RNAs that sedimented at 25S to 54S were a mixture of genome and anti-genome type. These results show that transcription and replication activities were associated with Sendai virus nucleocapsids obtained from infected cells and that some of the reaction products approached genome size.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-60-1-67
1982-05-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/jgv/60/1/JV0600010067.html?itemId=/content/journal/jgv/10.1099/0022-1317-60-1-67&mimeType=html&fmt=ahah

References

  1. Batt-Humphries S., Simonsen C., Ehrenfeld E. 1979; Full-length viral RNA synthesized in vitro by vesicular stomatitis virus-infected HeLa cell extracts. Virology 96:88–99
    [Google Scholar]
  2. Birnstiel M. L., Sells B. H., Purdom I. F. 1972; Kinetic complexity ofRNA molecules. Journal of Molecular Biology 63:21–39
    [Google Scholar]
  3. Blumberg B. M., Leppert M., Kolakofsky D. 1981; Interaction of VSV leader RNA and nucleocapsid protein may control VSV genome replication. Cell 23:837–845
    [Google Scholar]
  4. Breindl M., Holland J. J. 1976; Studies on in vitro transcription and translation of vesicular stomatitis virus mRNA. Virology 73:106–118
    [Google Scholar]
  5. Chinchar V. G., Portner A. 1981; Functions of Sendai virus nucleocapsid polypeptides: enzymatic activities in nucleocapsids following cleavage of polypeptide P by Staphylococcus aureus protease V8. Virology 109:59–71
    [Google Scholar]
  6. Choppin P. W., Compans R. W. 1975; Reproduction of paramyxoviruses. In Comprehensive Virology vol 4: pp 95–178 Edited by Fraenkel-Conrat H., Wagner R. R. New York: Plenum Press;
    [Google Scholar]
  7. Darlington R. W., Portner A., Kingsbury D. W. 1970; Sendai virus replication: an ultrastructural comparison of productive and abortive infections in avian cells. Journal of General Virology 9:169–177
    [Google Scholar]
  8. Emerson S. U. 1976; Vesicular stomatitis virus: structure and function of virion components. Current Topics in Microbiology and Immunology 73:1–34
    [Google Scholar]
  9. Friedman R. M., Metz D. H., Esteban R., Tovell D. R., Ball L. A., Kell L. M. 1972; Mechanism of interferon action: inhibition of viral messenger ribonucleic acid translation in L-cell extract. Journal of Virology 10:1184–1198
    [Google Scholar]
  10. Guild G. M., Stollar V. 1977; Defective interfering particles of Sindbis virus. V. Sequence relationships between SVSTD 42S RNA and intracellular defective viral RNAs. Virology 77:175–188
    [Google Scholar]
  11. Kingsbury D. W. 1974; The molecular biology of paramyxoviruses. Medical Microbiology and Immunology 160:73–83
    [Google Scholar]
  12. Kingsbury D. W. 1977; Paramyxoviruses. In The Molecular Biology of Animal Viruses vol 1: pp 349–382 Edited by Nayak D. P. New York: Marcel Dekker;
    [Google Scholar]
  13. Kolakofsky D., Bruschi A. 1975; Antigenomes in Sendai virions and Sendai virus-infected cells. Virology 66:185–191
    [Google Scholar]
  14. Lamb R. A., Choppin P. W. 1978; Determination by peptide mapping of the unique polypeptides in Sendai virions and infected cells. Virology 84:469–478
    [Google Scholar]
  15. Lehrach H., Diamond D., Wozney J. M., Boedtker H. 1977; RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry 16:4743–4751
    [Google Scholar]
  16. McMaster G. K., Carmichael G. C. 1977; Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proceedings of the National Academy of Sciences of the United States of America 74:4835–4838
    [Google Scholar]
  17. Marx P. A., Portner A., Kingsbury D. W. 1974; Sendai virion transcriptase complex: polypeptide composition and inhibition by virion envelope proteins. Journal of Virology 13:107–112
    [Google Scholar]
  18. Miller T. J., Stone H. O. 1977; Improved conditions for the in vitro synthesis of Newcastle disease virus messenger RNA. Abstract of the Annual Meeting of the American Society for Microbiology p 350
    [Google Scholar]
  19. Portner A. 1977; Association of nucleocapsid polypeptides with defective RNA synthesis in a temperature-sensitive mutant of Sendai virus. Virology 77:481–489
    [Google Scholar]
  20. Portner A., Kingsbury D. W. 1972; Identification of transcriptive and replicative intermediates in Sendai virus-infected cells. Virology 47:711–725
    [Google Scholar]
  21. Stone H. O., Portner A., Kingsbury D. W. 1971; Ribonucleic acid transcriptase in Sendai virions and infected cells. Virology 8:174–180
    [Google Scholar]
  22. Stone H. O., Kingsbury D. W., Darlington R. W. 1972; Sendai virus-induced transcriptase from infected ceil: polypeptides in the transcriptive complex. Journal of Virology 10:1037–1043
    [Google Scholar]
  23. Testa D., Chanda P. K., Banerjee A. K. 1980; In vitro synthesis of the full-length complement of the negative-strand genome RNA of vesicular stomatitis virus. Proceedings of the National Academy of Sciences of the United States of America 77:294–298
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-60-1-67
Loading
/content/journal/jgv/10.1099/0022-1317-60-1-67
Loading

Data & Media loading...

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