Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1987 Nov;61(11):3381–3387. doi: 10.1128/jvi.61.11.3381-3387.1987

Characterization of in vitro transcription and transcriptional products of measles virus.

J Ray 1, R S Fujinami 1
PMCID: PMC255932  PMID: 3669151

Abstract

Ribonucleoprotein complexes isolated from measles virus-infected HeLa cells contained an RNA-dependent RNA polymerase activity that catalyzed the incorporation of ribonucleotides into ribonucleic acid. The ribonucleoprotein complexes were composed of measles virus nucleoprotein, phosphoprotein, and a large protein, as well as viral RNA. The kinetics of RNA synthesis at different temperatures, time intervals, and protein, ribonucleotide, and mono- and divalent cation concentrations were analyzed. Enzyme activity was maximum at 4 h at 25 degrees C in the presence of 100 mM Na+-2.5 mM Mg2+-1 mM ribonucleotides. Actinomycin D and alpha-amanitin had no effect on the enzyme activity. Addition of cytoplasmic extracts from uninfected HeLa cells to the reaction mixture did not increase the incorporation of ribonucleotides into RNA. The in vitro synthesized RNAs were characterize by slot blot analysis and quantitated by densitometer scanning. All mRNAs coding for the structural proteins of measles virus were synthesized. Nucleoprotein RNA was the most abundant species made, followed by phosphoprotein, hemagglutinin, fusion protein, matrix protein, and large-protein RNAs. The system described here resulted in the first efficient transcription of measles virus RNA and analysis of products.

Full text

PDF
3385

Images in this article

3382Image
on p.3382
3384Image
on p.3384
3385Image
on p.3385

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aaslestad H. G., Clark H. F., Bishop D. H., Koprowski H. Comparison of the ribonucleic acid polymerases of two rhabdoviruses, Kern Canyon virus and vesicular stomatitis virus. J Virol. 1971 Jun;7(6):726–735. doi: 10.1128/jvi.7.6.726-735.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Abraham G., Banerjee A. K. Sequential transcription of the genes of vesicular stomatitis virus. Proc Natl Acad Sci U S A. 1976 May;73(5):1504–1508. doi: 10.1073/pnas.73.5.1504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baczko K., Carter M. J., Billeter M., ter Meulen V. Measles virus gene expression in subacute sclerosing panencephalitis. Virus Res. 1984 Oct;1(7):585–595. doi: 10.1016/0168-1702(84)90015-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baczko K., Liebert U. G., Billeter M., Cattaneo R., Budka H., ter Meulen V. Expression of defective measles virus genes in brain tissues of patients with subacute sclerosing panencephalitis. J Virol. 1986 Aug;59(2):472–478. doi: 10.1128/jvi.59.2.472-478.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Baltimore D., Huang A. S., Stampfer M. Ribonucleic acid synthesis of vesicular stomatitis virus, II. An RNA polymerase in the virion. Proc Natl Acad Sci U S A. 1970 Jun;66(2):572–576. doi: 10.1073/pnas.66.2.572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bishop D. H. Complete transcription by the transcriptase of vesicular stomatitis virus. J Virol. 1971 Apr;7(4):486–490. doi: 10.1128/jvi.7.4.486-490.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bishop D. H., Roy P. Properties of the product synthesized by vesicular stomatitis virus particles. J Mol Biol. 1971 Jun 28;58(3):799–814. doi: 10.1016/0022-2836(71)90041-6. [DOI] [PubMed] [Google Scholar]
  8. Breindl M., Holland J. J. Studies on the in vitro transcription and translation of vesicular stomatitis virus mRNA. Virology. 1976 Aug;73(1):106–118. doi: 10.1016/0042-6822(76)90065-9. [DOI] [PubMed] [Google Scholar]
  9. Carroll A. R., Wagner R. R. Role of the membrane (M) protein in endogenous inhibition of in vitro transcription by vesicular stomatitis virus. J Virol. 1979 Jan;29(1):134–142. doi: 10.1128/jvi.29.1.134-142.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Combard A., Printz Ane C. Inhibition of vesicular stomatitis virus transcriptase complex by the virion envelope M protein. Biochem Biophys Res Commun. 1979 May 14;88(1):117–123. doi: 10.1016/0006-291x(79)91704-2. [DOI] [PubMed] [Google Scholar]
  11. Dowling P. C., Blumberg B. M., Kolakofsky D., Cook P., Jotkowitz A., Prineas J. W., Cook S. D. Measles virus nucleic acid sequences in human brain. Virus Res. 1986 Jul;5(1):97–107. doi: 10.1016/0168-1702(86)90068-7. [DOI] [PubMed] [Google Scholar]
  12. Dowling P. C., Blumberg B. M., Menonna J., Adamus J. E., Cook P., Crowley J. C., Kolakofsky D., Cook S. D. Transcriptional map of the measles virus genome. J Gen Virol. 1986 Sep;67(Pt 9):1987–1992. doi: 10.1099/0022-1317-67-9-1987. [DOI] [PubMed] [Google Scholar]
  13. Flamand A., Bishop D. H. Primary in vivo transcription of vesicular stomatitis virus and temperature-sensitive mutants of five vesicular stomatitis virus complementation groups. J Virol. 1973 Dec;12(6):1238–1252. doi: 10.1128/jvi.12.6.1238-1252.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fujinami R. S., Oldstone M. B. Alterations in expression of measles virus polypeptides by antibody: molecular events in antibody-induced antigenic modulation. J Immunol. 1980 Jul;125(1):78–85. [PubMed] [Google Scholar]
  15. Fujinami R. S., Sissons J. G., Oldstone M. B. Immune reactive measles virus polypeptides on the cell's surface: turnover and relationship of the glycoproteins to each other and to HLA determinants. J Immunol. 1981 Sep;127(3):935–940. [PubMed] [Google Scholar]
  16. Glazier K., Raghow R., Kingsbury D. W. Regulation of Sendai virus transcription: evidence for a single promoter in vivo. J Virol. 1977 Mar;21(3):863–871. doi: 10.1128/jvi.21.3.863-871.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Haase A. T., Ventura P., Gibbs C. J., Jr, Tourtellotte W. W. Measles virus nucleotide sequences: detection by hybridization in situ. Science. 1981 May 8;212(4495):672–675. doi: 10.1126/science.7221554. [DOI] [PubMed] [Google Scholar]
  18. Hill V. M., Harmon S. A., Summers D. F. Stimulation of vesicular stomatitis virus in vitro RNA synthesis by microtubule-associated proteins. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5410–5413. doi: 10.1073/pnas.83.15.5410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hill V. M., Summers D. F. Synthesis of VSV RNPs in vitro by cellular VSV RNPs added to uninfected HeLa cell extracts: VSV protein requirements for replication in vitro. Virology. 1982 Dec;123(2):407–419. doi: 10.1016/0042-6822(82)90273-2. [DOI] [PubMed] [Google Scholar]
  20. Horta-Barbosa L., Fuccillo D. A., Sever J. L., Zeman W. Subacute sclerosing panencephalitis: isolation of measles virus from a brain biopsy. Nature. 1969 Mar 8;221(5184):974–974. doi: 10.1038/221974a0. [DOI] [PubMed] [Google Scholar]
  21. Huang A. S., Baltimore D., Bratt M. A. Ribonucleic acid polymerase in virions of Newcastle disease virus: comparison with the vesicular stomatitis virus polymerase. J Virol. 1971 Mar;7(3):389–394. doi: 10.1128/jvi.7.3.389-394.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jashés M., Sandino A. M., Faúndez G., Avendaño L. F., Spencer E. In vitro transcription of human pararotavirus. J Virol. 1986 Jan;57(1):183–190. doi: 10.1128/jvi.57.1.183-190.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Johnson R. T., Griffin D. E., Hirsch R. L., Wolinsky J. S., Roedenbeck S., Lindo de Soriano I., Vaisberg A. Measles encephalomyelitis--clinical and immunologic studies. N Engl J Med. 1984 Jan 19;310(3):137–141. doi: 10.1056/NEJM198401193100301. [DOI] [PubMed] [Google Scholar]
  24. Mountcastle W. E., Choppin P. W. A comparison of the polypeptides of four measles virus strains. Virology. 1977 May 15;78(2):463–474. doi: 10.1016/0042-6822(77)90123-4. [DOI] [PubMed] [Google Scholar]
  25. Moyer S. A., Baker S. C., Lessard J. L. Tubulin: a factor necessary for the synthesis of both Sendai virus and vesicular stomatitis virus RNAs. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5405–5409. doi: 10.1073/pnas.83.15.5405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Payne F. E., Baublis J. V., Itabashi H. H. Isolation of measles virus from cell cultures of brain from a patient with subacute sclerosing panencephalitis. N Engl J Med. 1969 Sep 11;281(11):585–589. doi: 10.1056/NEJM196909112811103. [DOI] [PubMed] [Google Scholar]
  27. Perrault J., Kingsbury D. T. Inhibitor of vesicular stomatitis virus transcriptase in purified virions. Nature. 1974 Mar 1;248(5443):45–47. doi: 10.1038/248045a0. [DOI] [PubMed] [Google Scholar]
  28. Rima B. K., Baczko K., Clarke D. K., Curran M. D., Martin S. J., Billeter M. A., ter Meulen V. Characterization of clones for the sixth (L) gene and a transcriptional map for morbilliviruses. J Gen Virol. 1986 Sep;67(Pt 9):1971–1978. doi: 10.1099/0022-1317-67-9-1971. [DOI] [PubMed] [Google Scholar]
  29. Robinson W. S. Ribonucleic acid polymerase activity in Sendai virions and nucleocapsid. J Virol. 1971 Jul;8(1):81–86. doi: 10.1128/jvi.8.1.81-86.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Seifried A. S., Albrecht P., Milstien J. B. Characterization of an RNA-dependent RNA polymerase activity associated with measles virus. J Virol. 1978 Mar;25(3):781–787. doi: 10.1128/jvi.25.3.781-787.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Shioda T., Iwasaki K., Shibuta H. Determination of the complete nucleotide sequence of the Sendai virus genome RNA and the predicted amino acid sequences of the F, HN and L proteins. Nucleic Acids Res. 1986 Feb 25;14(4):1545–1563. doi: 10.1093/nar/14.4.1545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Stallcup K. C., Wechsler S. L., Fields B. N. Purification of measles virus and characterization of subviral components. J Virol. 1979 Apr;30(1):166–176. doi: 10.1128/jvi.30.1.166-176.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Stone H. O., Portner A., Kingsbury D. W. Ribonucleic acid transcriptases in Sendai Virions and infected cells. J Virol. 1971 Aug;8(2):174–180. doi: 10.1128/jvi.8.2.174-180.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wechsler S. L., Fields B. N. Intracellular synthesis of measles virus-specified polypeptides. J Virol. 1978 Jan;25(1):285–297. doi: 10.1128/jvi.25.1.285-297.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES