1887

Abstract

SUMMARY

The kinetics of H-uridine incorporation into measles-infected Vero cells demonstrated that maximum virus-specific RNA synthesis occurred between 16 and 20 h after infection. Sedimentation analysis on sucrose gradients revealed the presence of four species of RNA having sedimentation coefficients of 4S, 12 to 26S, 28 to 36S, and 50S. Annealing studies showed that RNA sedimenting in the 12 to 36S regions was 100% complementary in base sequence to nucleocapsid 50S RNA, and at least 96% of the 50S genomic RNA was transcribed during virus replication. Polynucleotide binding experiments and ribonuclease treatment indicated that poly(A) sequences were associated with the intracellular 12 to 26S, 28 to 36S and 50S RNAs. Denaturation of intracellular 50S RNA followed by sucrose gradient centrifugation demonstrated that this was a mixture of genomic 50S and heterogeneous RNAs which sedimented at 4 to 40S. The genomic RNA did not contain poly(A) sequences, and these are presumably associated with the heterogeneously sedimenting RNAs. The size of poly(A) sequences present on the 12 to 36S RNAs was estimated to be in the range of 70 to 140 nucleotides. Treatment of the 12 to 36S RNAs and their poly(A) sequences with polynucleotide phosphorylase indicated that the poly(A) was located on the 3′ end of the RNAs, but that under the experimental conditions used this was protected by the secondary structure of the molecules.

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1977-06-01
2024-10-04
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References

  1. Banerjee A. K., Moyer S. A., Rhodes D. P. 1974; Studies on the in vitro adenylation of RNA by vesicular stomatitis virus. Virology 61:547–558
    [Google Scholar]
  2. Bratt M. A., Collins B. S., Hightower L. L., Kaplan J., Tsipis J. E., Weiss S. R. 1975; Transcription and translation of Newcastle disease virus RNA. In Negative Strand Viruses p 387 Edited by Mahy B. W. J., Barry R. D. New York and London: Academic Press;
    [Google Scholar]
  3. Bratt M. A., Robinson W. S. 1967; Ribonucleic acid synthesis in cells infected with Newcastle disease virus. Journal of Molecular Biology 23:1–21
    [Google Scholar]
  4. Bratt M. A., Robinson W. S. 1971; Evidence for an RNA replicative intermediate in cells infected with Newcastle disease virus. Journal of General Virology 10:139–145
    [Google Scholar]
  5. Carter C., Schluederberg A., Black F. L. 1973; Viral RNA synthesis in measles-infected cells. Virology S3:379–383
    [Google Scholar]
  6. Compans R. W., Choppin P. W. 1967; Isolation and properties of the helical nucleocapsid of the parainfluenza virus SV5. Proceedings of the National Academy of Sciences of the United States of America 57:949–956
    [Google Scholar]
  7. East J. L., Kingsbury D. W. 1971; Mumps virus replication in chick embryo lung cells: properties of ribonucleic acids in virions and infected cells. Journal of Virology 8:161–173
    [Google Scholar]
  8. Gorski J., Morrison M. R., Merkel C. G., Lingrel J. B. 1974; Size heterogeneity of polyadenylate sequences in mouse globin messenger RNA. Journal of Molecular Biology 86:363–371
    [Google Scholar]
  9. Hall W. W., Martin S. J. 1973; Purification and characterization of measles virus. Journal of General Virology 19:175–188
    [Google Scholar]
  10. Hall W. W., Martin S. J., Gould E. A. 1974; Defective interfering particles produced during the replication of measles virus. Medical Microbiology and Immunology 160:155–164
    [Google Scholar]
  11. Kiley M. P., Payne F. E. 1974; Replication of measles virus: continued synthesis of nucleocapsid RNA and increased synthesis of mRNA in the presence of cycloheximide. Journal of Virology 14:758–764
    [Google Scholar]
  12. Kingsbury D. W. 1972; Paramyxovirus replication. Current Topics in Microbiology and Immunology 59:1–33
    [Google Scholar]
  13. Kingsbury D. W. 1973; Cell-free translation of paramyxovirus messenger RNA. Journal of Virology 12:1020–1027
    [Google Scholar]
  14. Kingsbury D. W. 1974; The molecular biology of the paramyxoviruses. Medical Microbiology and Immunology 160:73–83
    [Google Scholar]
  15. Kolakofsky D., Bruschi A. 1975; Antigenomes in Sendai virions and Sendai virus-infected cells. Virology 66:185–199
    [Google Scholar]
  16. Lee S. Y., Mendecki J., Brawermann G. 1971; A polynucleotide segment rich in adenylic acid in the rapidly labelled polyribosomal RNA component of mouse sarcoma 180 ascites cells. Proceedings of the National Academy of Sciences of the United States of America 68:1331–1335
    [Google Scholar]
  17. Lindberg U., Persson T. 1972; Isolation of an RNA from KB cells by affinity chromatography of polyuridylic acid covalently linked to Sepharose. European Journal of Biochemistry 31:246–254
    [Google Scholar]
  18. Marshall S., Gillespie D. 1972; Poly U tracts absent from viral RNA. Nature New Biology 240:43–45
    [Google Scholar]
  19. Marx P. A. Jun, Pridgen C., Kingsbury D. W. 1975; Location and abundance of poly(A) sequences in Sendai virus RNA molecules. Journal of General Virology 27:247–250
    [Google Scholar]
  20. Moyer S. A., Grubman M. J., Ehrenfeld E., Banerjee A. K. 1975; Studies on the in vivo and in vitro messenger RNA species of vesicular stomatitis virus. Virology 67:463–473
    [Google Scholar]
  21. Portner A., Bussel R. H. 1973; Measles virus ribonucleic and protein synthesis: effects of 6-azauridine and cycloheximide on viral replication. Journal of Virology x1:46–53
    [Google Scholar]
  22. Pridgen C., Kingsbury D. W. 1972; Adenylate-rich sequences in Sendai virus transcripts from infected cells. Journal of Virology 10:314–317
    [Google Scholar]
  23. Schluederberg A. 1971; Measles virus RNA. Biochemical and Biophysical Research Communications 42:1012–1015
    [Google Scholar]
  24. Sheiness P., Darnell J. E. 1973; Polyadenylic acid segment in mRNA becomes shorter with age. Nature New Biology 241:265–268
    [Google Scholar]
  25. Sheldon R., Kates J., Kelley D. E., Perry R. P. 1972; Polyadenylic acid sequences on the 3′ termini of vaccinia messenger ribonucleic acid and mannalian nuclear and messenger ribonucleic acid. Biochemistry II:3829–3834
    [Google Scholar]
  26. Weiss S. R., Bratt M. A. 1974; Polyadenylate sequences on Newcastle disease virus mRNA synthesized in vivo and in vitro. Journal of Virology 13:1220–1230
    [Google Scholar]
  27. Winston S. H., Rustigian R., Bratt M. A. 1973; Persistent infection of cells in culture by measles virus. 3. Comparison of virus-specific RNA synthesized in primary and persistent infection in HeLa cells. Journal of Virology 11:926–932
    [Google Scholar]
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