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. 1970 Sep;6(3):259–268. doi: 10.1128/jvi.6.3.259-268.1970

In Vitro Product of a Ribonucleic Acid Polymerase Induced by Influenza Virus

B W J Mahy 1, P A Bromley 1
PMCID: PMC376118  PMID: 5480408

Abstract

The ribonucleic acid (RNA)-dependent RNA polymerase induced in the microsomal fraction of cells infected with influenza virus synthesized a mixture of single-and double-stranded RNA in vitro. The single-stranded RNA sedimented mainly in the 8S region on sucrose density gradients, with a smaller proportion of the RNA sedimenting at 18S. This sedimentation pattern corresponds closely to that of incomplete influenza virus RNA. The double-stranded RNA formed in vitro sedimented at 11S, but molecules which may be replicative intermediate, sedimenting at 14 to 20S, were also detected in the in vitro reaction product. Similar species of RNA were detected in vivo by pulse-labeling infected cells at the time of polymerase harvest, but the proportion of each RNA species was different, most of the RNA being single-stranded and sedimenting in the 18S region. An 11S double-stranded RNA was also synthesized in vivo. Pulse chase analysis of the double-stranded RNA synthesized in vitro showed that most is stable, and only a small proportion turns over during the reaction. A proportion of the RNA formed in vitro could be annealed to RNA formed in infected cells and to RNA extracted from purified virus.

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Selected References

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

  1. Arlinghaus R. B., Polatnick J. In vitro products of a membrane-free foot-and-mouth disease virus ribonucleic acid polymerase. Virology. 1969 Feb;37(2):252–261. doi: 10.1016/0042-6822(69)90205-0. [DOI] [PubMed] [Google Scholar]
  2. Arlinghaus R. B., Polatnick J. The isolation of two enzyme-ribonucleic acid complexes involved in the synthesis of foot-and-mouth disease virus ribonucleic acid. Proc Natl Acad Sci U S A. 1969 Mar;62(3):821–828. doi: 10.1073/pnas.62.3.821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BARRY R. D., IVES D. R., CRUICKSHANK J. G. Participation of deoxyribonucleic acid in the multiplication of influenza virus. Nature. 1962 Jun 23;194:1139–1140. doi: 10.1038/1941139a0. [DOI] [PubMed] [Google Scholar]
  4. BARRY R. D. THE EFFECTS OF ACTINOMYCIN D AND ULTRAVIOLET IRRADIATION ON THE PRODUCTION OF FOWL PLAGUE VIRUS. Virology. 1964 Dec;24:563–569. doi: 10.1016/0042-6822(64)90208-9. [DOI] [PubMed] [Google Scholar]
  5. Baltimore D., Girard M. An intermediate in the synthesis of poliovirus RNA. Proc Natl Acad Sci U S A. 1966 Aug;56(2):741–748. doi: 10.1073/pnas.56.2.741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Baltimore D. Structure of the poliovirus replicative intermediate RNA. J Mol Biol. 1968 Mar 14;32(2):359–368. doi: 10.1016/0022-2836(68)90015-6. [DOI] [PubMed] [Google Scholar]
  7. Barry R. D., Davies P. The sedimentation of influenza virus and its RNA in sucrose density gradients. J Gen Virol. 1968 Jan;2(1):59–69. doi: 10.1099/0022-1317-2-1-59. [DOI] [PubMed] [Google Scholar]
  8. Billeter M. A., Weissmann C., Warner R. C. Replication of viral ribonucleic acid. IX. Properties of double-stranded RNA from Escherichia coli infected with bacteriophage MS2. J Mol Biol. 1966 May;17(1):145–173. doi: 10.1016/s0022-2836(66)80101-8. [DOI] [PubMed] [Google Scholar]
  9. Borland R., Mahy B. W. Deoxyribonucleic acid-dependent ribonucleic acid polymerase activity in cells infected with influenza virus. J Virol. 1968 Jan;2(1):33–39. doi: 10.1128/jvi.2.1.33-39.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Britten R. J., Roberts R. B. High-Resolution Density Gradient Sedimentation Analysis. Science. 1960 Jan 1;131(3392):32–33. doi: 10.1126/science.131.3392.32. [DOI] [PubMed] [Google Scholar]
  11. Davies P., Barry R. D. Nucleic acid of influenza virus. Nature. 1966 Jul 23;211(5047):384–387. doi: 10.1038/211384a0. [DOI] [PubMed] [Google Scholar]
  12. Deusberg P. H., Robinson W. S. On the structure and replication of influenza virus. J Mol Biol. 1967 May 14;25(3):383–405. doi: 10.1016/0022-2836(67)90193-3. [DOI] [PubMed] [Google Scholar]
  13. Duesberg P. H. The RNA of influenza virus. Proc Natl Acad Sci U S A. 1968 Mar;59(3):930–937. doi: 10.1073/pnas.59.3.930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Feix G., Pollet R., Weissmann C. Replication of viral RNA, XVI. Enzymatic synthesis of infectious virual RNA with noninfectious Q-beta minus strands as template. Proc Natl Acad Sci U S A. 1968 Jan;59(1):145–152. doi: 10.1073/pnas.59.1.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. GEIDUSCHEK E. P., MOOHR J. W., WEISS S. B. The secondary structure of complementary RNA. Proc Natl Acad Sci U S A. 1962 Jun 15;48:1078–1086. doi: 10.1073/pnas.48.6.1078. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Girard M. In vitro synthesis of poliovirus ribonucleic acid: role of the replicative intermediate. J Virol. 1969 Apr;3(4):376–384. doi: 10.1128/jvi.3.4.376-384.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. HIRST G. K. Genetic recombination with Newcastle disease virus, polioviruses, and influenza. Cold Spring Harb Symp Quant Biol. 1962;27:303–309. doi: 10.1101/sqb.1962.027.001.028. [DOI] [PubMed] [Google Scholar]
  19. Ho P. P., Walters C. P. Influenza virus-induced ribonucleic acid nucleotidyltransferase and the effect of actinomycin D on its formation. Biochemistry. 1966 Jan;5(1):231–235. doi: 10.1021/bi00865a030. [DOI] [PubMed] [Google Scholar]
  20. KIRBY K. S. ISOLATION AND CHARACTERIZATION OF RIBOSOMAL RIBONUCLEIC ACID. Biochem J. 1965 Jul;96:266–269. doi: 10.1042/bj0960266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Lodish H. F., Zinder N. D. Replication of the RNA of Bacteriophage f2. Science. 1966 Apr 15;152(3720):372–377. doi: 10.1126/science.152.3720.372. [DOI] [PubMed] [Google Scholar]
  23. Mahy B. W., Bromley P. A. Synthesis of ribonuclease-resistant ribonucleic acid by fowl-plague-virus-induced ribonucleic acid polymerase. Biochem J. 1969 Oct;114(4):64P–64P. doi: 10.1042/bj1140064pa. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nayak D. P., Baluda M. A. An intermediate in the replication of influenza virus RNA. Proc Natl Acad Sci U S A. 1968 Jan;59(1):184–191. doi: 10.1073/pnas.59.1.184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nayak D. P., Baluda M. A. Characterization of influenza virus ribonucleic acid duplex produced by annealing in vitro. J Virol. 1969 Mar;3(3):318–325. doi: 10.1128/jvi.3.3.318-325.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nayak D. P., Baluda M. A. Isolation and partial characterization of nucleic acid of influenza virus. J Virol. 1967 Dec;1(6):1217–1223. doi: 10.1128/jvi.1.6.1217-1223.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nayak D. P., Baluda M. A. Ribonucleic acid synthesis in cells infected with influenza virus. J Virol. 1968 Feb;2(2):99–109. doi: 10.1128/jvi.2.2.99-109.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nayak D. P. Influenza virus: structure, replication and defectiveness. Fed Proc. 1969 Nov-Dec;28(6):1858–1866. [PubMed] [Google Scholar]
  29. Pace N. R., Bishop D. H., Spiegelman S. Examination of the Qbeta-replicase reaction by sucrose gradient and gel electrophoresis. J Virol. 1967 Aug;1(4):771–778. doi: 10.1128/jvi.1.4.771-778.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Plagemann P. G. Choline metabolism and membrane formation in rat hepatoma cells grown in suspension culture. I. Incorporation of choline into phosphatidylcholine of mitochondria and other membranous structures and effect of metabolic inhibitors. Arch Biochem Biophys. 1968 Oct;128(1):70–87. doi: 10.1016/0003-9861(68)90009-x. [DOI] [PubMed] [Google Scholar]
  31. Plagemann P. G., Swim H. E. Replication of mengovirus. II. General properties of the viral-induced ribonucleic acid polymerase. J Bacteriol. 1966 Jun;91(6):2327–2332. doi: 10.1128/jb.91.6.2327-2332.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Plagemann P. G., Swim H. E. Synthesis of ribonucleic acid by mengovirus-induced RNA polymerase in vitro: nature of products and of RNase-resistant intermediate. J Mol Biol. 1968 Jul 14;35(1):13–25. doi: 10.1016/s0022-2836(68)80034-8. [DOI] [PubMed] [Google Scholar]
  33. Pons M. W., Hirst G. K. Polyacrylamide gel electrophoresis of influenza virus RNA. Virology. 1968 Feb;34(2):385–388. doi: 10.1016/0042-6822(68)90257-2. [DOI] [PubMed] [Google Scholar]
  34. Pons M. W. Some characteristics of double-strandedinfluenza virus ribonucleic acid. Arch Gesamte Virusforsch. 1967;22(1):203–209. doi: 10.1007/BF01240514. [DOI] [PubMed] [Google Scholar]
  35. Pons M., Hirst G. K. The single- and double-stranded RNA's and the proteins of incomplete influenza virus. Virology. 1969 May;38(1):68–72. doi: 10.1016/0042-6822(69)90128-7. [DOI] [PubMed] [Google Scholar]
  36. REICH E., FRANKLIN R. M., SHATKIN A. J., TATUMEL Action of actinomycin D on animal cells and viruses. Proc Natl Acad Sci U S A. 1962 Jul 15;48:1238–1245. doi: 10.1073/pnas.48.7.1238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Ruck B. J., Brammer K. W., Page M. G., Coombes J. D. The detection and characterization of an induced RNA polymerase in the chorioallantoic membranes of embryonated eggs infected with influenza A2 virus. Virology. 1969 Sep;39(1):31–41. doi: 10.1016/0042-6822(69)90345-6. [DOI] [PubMed] [Google Scholar]
  38. Scholtissek C., Rott R. Ribonucleic acid nucleotidyl transferase induced in chick fibroblasts after infection with an influenza virus. J Gen Virol. 1969 Jan;4(1):125–137. doi: 10.1099/0022-1317-4-1-125. [DOI] [PubMed] [Google Scholar]
  39. Scholtissek C. Synthesis in vitro of RNA complementary to parental viral RNA by RNA polymerase induced by influenza virus. Biochim Biophys Acta. 1969 Apr 22;179(2):389–397. doi: 10.1016/0005-2787(69)90047-1. [DOI] [PubMed] [Google Scholar]
  40. Skehel J. J., Burke D. C. Ribonucleic acid synthesis in chick embryo cells infected with fowl-plague virus. J Virol. 1969 Apr;3(4):429–438. doi: 10.1128/jvi.3.4.429-438.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Sreevalsan T., Yin F. H. Sindbis virus-induced viral ribonucleic acid polymerase. J Virol. 1969 Jun;3(6):599–604. doi: 10.1128/jvi.3.6.599-604.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Tershak D. R. The in vitro product of poliovirus polymerase. Can J Microbiol. 1969 Feb;15(2):240–242. doi: 10.1139/m69-042. [DOI] [PubMed] [Google Scholar]
  43. Weissmann C., Ochoa S. Replication of phage RNA. Prog Nucleic Acid Res Mol Biol. 1967;6:353–399. doi: 10.1016/s0079-6603(08)60530-9. [DOI] [PubMed] [Google Scholar]
  44. White D. O., Cheyne I. M. Early events in the eclipse phase of influenza and parainfluenza virus infection. Virology. 1966 May;29(1):49–59. doi: 10.1016/0042-6822(66)90195-4. [DOI] [PubMed] [Google Scholar]

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