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. 1970 Jul;6(1):1–11. doi: 10.1128/jvi.6.1.1-11.1970

Transcription In Vitro by Reovirus-Associated Ribonucleic Acid-Dependent Polymerase 1

A K Banerjee 1, A J Shatkin 1
PMCID: PMC376083  PMID: 5529847

Abstract

Digestion of purified reovirus type 3 with chymotrypsin degrades 70% of the viral protein and converts the virions to subviral particles (SVP). The SVP contain 3 of the 6 viral structural proteins and all 10 double-stranded ribonucleic acid (RNA) genome segments but not adenine-rich, single-stranded RNA. An RNA polymerase which is structurally associated with SVP transcribes one strand of each genome segment by a conservative mechanism in vitro. The single-stranded products include large (1.2 × 106 daltons), medium (0.7 × 106 daltons), and small (0.4 × 106 daltons) molecules which hybridize exclusively with the corresponding genome segments. The enzyme obtained by heating virions at 60 C synthesizes similar products. Kinetic and pulse-chase studies indicate that the different-sized products are synthesized simultaneously but at rates which are in the order: small > medium > large.

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Selected References

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

  1. Bellamy A. R., Joklik W. K. Studies on reovirus RNA. II. Characterization of reovirus messenger RNA and of the genome RNA segments from which it is transcribed. J Mol Biol. 1967 Oct 14;29(1):19–26. doi: 10.1016/0022-2836(67)90178-7. [DOI] [PubMed] [Google Scholar]
  2. Bellamy A. R., Joklik W. K. Studies on the A-rich RNA of reovirus. Proc Natl Acad Sci U S A. 1967 Oct;58(4):1389–1395. doi: 10.1073/pnas.58.4.1389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bellamy A. R., Shapiro L., August J. T., Joklik W. K. Studies on reovirus RNA. I. Characterization of reovirus genome RNA. J Mol Biol. 1967 Oct 14;29(1):1–17. doi: 10.1016/0022-2836(67)90177-5. [DOI] [PubMed] [Google Scholar]
  4. Borsa J., Graham A. F. Reovirus: RNA polymerase activity in purified virions. Biochem Biophys Res Commun. 1968 Dec 30;33(6):895–901. doi: 10.1016/0006-291x(68)90396-3. [DOI] [PubMed] [Google Scholar]
  5. Boyer P. D. The inhibition of pyruvate kinase by ATP: a Mg++ buffer system for use in enzyme studies. Biochem Biophys Res Commun. 1969 Mar 10;34(5):702–706. doi: 10.1016/0006-291x(69)90795-5. [DOI] [PubMed] [Google Scholar]
  6. Burgess R. R., Travers A. A., Dunn J. J., Bautz E. K. Factor stimulating transcription by RNA polymerase. Nature. 1969 Jan 4;221(5175):43–46. doi: 10.1038/221043a0. [DOI] [PubMed] [Google Scholar]
  7. FRAENKEL-CONRAT H., SINGER B., TSUGITA A. Purification of viral RNA by means of bentonite. Virology. 1961 May;14:54–58. doi: 10.1016/0042-6822(61)90131-3. [DOI] [PubMed] [Google Scholar]
  8. GOMATOS P. J., TAMM I., DALES S., FRANKLIN R. M. Reovirus type 3: physical characteristics and interaction with L cells. Virology. 1962 Jul;17:441–454. doi: 10.1016/0042-6822(62)90139-3. [DOI] [PubMed] [Google Scholar]
  9. Gomatos P. J. RNA synthesis in reovirus-infected L929 mouse fibroblasts. Proc Natl Acad Sci U S A. 1967 Oct;58(4):1798–1805. doi: 10.1073/pnas.58.4.1798. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gomatos P. J., Tamm I. THE SECONDARY STRUCTURE OF REOVIRUS RNA. Proc Natl Acad Sci U S A. 1963 May;49(5):707–714. doi: 10.1073/pnas.49.5.707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kates J. R., McAuslan B. R. Poxvirus DNA-dependent RNA polymerase. Proc Natl Acad Sci U S A. 1967 Jul;58(1):134–141. doi: 10.1073/pnas.58.1.134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Krug R. M., Gomatos P. J. Absence of adenine-rich ribonucleic acid from purified infectious reovirus 3. J Virol. 1969 Nov;4(5):642–650. doi: 10.1128/jvi.4.5.642-650.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kudo H., Graham A. F. Selective inhibition of reovirus induced RNA in L cells. Biochem Biophys Res Commun. 1966 Jul 20;24(2):150–155. doi: 10.1016/0006-291x(66)90711-x. [DOI] [PubMed] [Google Scholar]
  14. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  15. Lewandowski L. J., Kalmakoff J., Tanada Y. Characterization of a Ribonucleic Acid Polymerase Activity Associated with Purified Cytoplasmic Polyhedrosis Virus of the Silkworm Bombyx mori. J Virol. 1969 Dec;4(6):857–865. doi: 10.1128/jvi.4.6.857-865.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Loening U. E. The fractionation of high-molecular-weight ribonucleic acid by polyacrylamide-gel electrophoresis. Biochem J. 1967 Jan;102(1):251–257. doi: 10.1042/bj1020251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Loh P. C., Shatkin A. J. Structural proteins of reoviruses. J Virol. 1968 Nov;2(11):1353–1359. doi: 10.1128/jvi.2.11.1353-1359.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. MAYOR H. D., JAMISON R. M., JORDAN L. E., VANMITCHELL M. REOVIRUSES. II. STRUCTURE AND COMPOSITION OF THE VIRION. J Bacteriol. 1965 Jun;89:1548–1556. doi: 10.1128/jb.89.6.1548-1556.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Maitra U., Barash F. DNA-dependent synthesis of RNA by Escherichia coli RNA polymerase: release and reinitiation of RNA chains from DNA templates. Proc Natl Acad Sci U S A. 1969 Oct;64(2):779–786. doi: 10.1073/pnas.64.2.779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Munyon W., Paoletti E., Grace J. T., Jr RNA polymerase activity in purified infectious vaccinia virus. Proc Natl Acad Sci U S A. 1967 Dec;58(6):2280–2287. doi: 10.1073/pnas.58.6.2280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Reich E., Goldberg I. H. Actinomycin and nucleic acid function. Prog Nucleic Acid Res Mol Biol. 1964;3:183–234. doi: 10.1016/s0079-6603(08)60742-4. [DOI] [PubMed] [Google Scholar]
  22. Roberts J. W. Termination factor for RNA synthesis. Nature. 1969 Dec 20;224(5225):1168–1174. doi: 10.1038/2241168a0. [DOI] [PubMed] [Google Scholar]
  23. SEBRING E. D., SALZMAN N. P. AN IMPROVED PROCEDURE FOR MEASURING THE DISTRIBUTION OF P32O4--AMONG THE NUCLEOTIDES OF RIBONUCLEIC ACID. Anal Biochem. 1964 May;8:126–129. doi: 10.1016/0003-2697(64)90177-0. [DOI] [PubMed] [Google Scholar]
  24. Shatkin A. J. Inactivity of purified reovirus RNA as a template for E. coli polymerases in vitro. Proc Natl Acad Sci U S A. 1965 Dec;54(6):1721–1728. doi: 10.1073/pnas.54.6.1721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Shatkin A. J., Rada B. Reovirus-directed ribonucleic acid synthesis in infected L cells. J Virol. 1967 Feb;1(1):24–35. doi: 10.1128/jvi.1.1.24-35.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Shatkin A. J., Sipe J. D., Loh P. Separation of ten reovirus genome segments by polyacrylamide gel electrophoresis. J Virol. 1968 Oct;2(10):986–991. doi: 10.1128/jvi.2.10.986-991.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Shatkin A. J., Sipe J. D. RNA polymerase activity in purified reoviruses. Proc Natl Acad Sci U S A. 1968 Dec;61(4):1462–1469. doi: 10.1073/pnas.61.4.1462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Shatkin A. J., Sipe J. D. Single-stranded, adenine-rich RNA from purified reoviruses. Proc Natl Acad Sci U S A. 1968 Jan;59(1):246–253. doi: 10.1073/pnas.59.1.246. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sippel A., Hartmann G. Mode of action of rafamycin on the RNA polymerase reaction. Biochim Biophys Acta. 1968 Mar 18;157(1):218–219. doi: 10.1016/0005-2787(68)90286-4. [DOI] [PubMed] [Google Scholar]
  30. Skehel J. J., Joklik W. K. Studies on the in vitro transcription of reovirus RNA catalyzed by reovirus cores. Virology. 1969 Dec;39(4):822–831. doi: 10.1016/0042-6822(69)90019-1. [DOI] [PubMed] [Google Scholar]
  31. Smith R. E., Zweerink H. J., Joklik W. K. Polypeptide components of virions, top component and cores of reovirus type 3. Virology. 1969 Dec;39(4):791–810. doi: 10.1016/0042-6822(69)90017-8. [DOI] [PubMed] [Google Scholar]
  32. So A. G., Davie E. W., Epstein R., Tissières A. Effects of cations on DNA-dependent RNA polymerase. Proc Natl Acad Sci U S A. 1967 Oct;58(4):1739–1746. doi: 10.1073/pnas.58.4.1739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Stirpe F., Fiume L. Studies on the pathogenesis of liver necrosis by alpha-amanitin. Effect of alpha-amanitin on ribonucleic acid synthesis and on ribonucleic acid polymerase in mouse liver nuclei. Biochem J. 1967 Nov;105(2):779–782. doi: 10.1042/bj1050779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. VASQUEZ C., TOURNIER P. The morphology of reovirus. Virology. 1962 Aug;17:503–510. doi: 10.1016/0042-6822(62)90149-6. [DOI] [PubMed] [Google Scholar]
  35. Watanabe Y., Graham A. F. Structural units of reovirus ribonucleic acid and their possible functional significance. J Virol. 1967 Aug;1(4):665–677. doi: 10.1128/jvi.1.4.665-677.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Watanabe Y., Kudo H., Graham A. F. Selective inhibition of reovirus ribonucleic acid synthesis by cycloheximide. J Virol. 1967 Feb;1(1):36–44. doi: 10.1128/jvi.1.1.36-44.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Watanabe Y., Millward S., Graham A. F. Regulation of transcription of the Reovirus genome. J Mol Biol. 1968 Aug 28;36(1):107–123. doi: 10.1016/0022-2836(68)90223-4. [DOI] [PubMed] [Google Scholar]
  38. Watanabe Y., Prevec L., Graham A. F. Specificity in transcription of the reovirus genome. Proc Natl Acad Sci U S A. 1967 Sep;58(3):1040–1046. doi: 10.1073/pnas.58.3.1040. [DOI] [PMC free article] [PubMed] [Google Scholar]

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