Abstract
Influenza viral mRNA, i.e., complementary RNA (cRNA), isolated from infected cells , was resolved into six different species by electrophoresis in 2.1% acrylamide gels containing 6 M urea. The cRNA's were grouped into three size classes: L (large), M (medium-size), and S (small). Similarly, when gels were sliced for analysis, the virion RNA (vRNA) also distributed into six peaks because the three largest vRNA segments were closely spaced and were resolved only when the gels were autoradiographed or stained. Because of their attached polyadenylic acid [poly(A)]sequences, the cRNA segments migrated more slowly than did the corresponding vRNA segments during gel electrophoresis. After removal of the poly(A) by RNase H, the cRNA and vRNA segments comigrated, indicating that they were approximately the same size. One of the cRNA segments, S2, was shown by annealing to contain the genetic information in the vRNA segment with which it comigrated, strongly suggesting that each cRNA segment was transcribed from the vRNA segment of the same size. In contrast to the vRNA segments, which when isolated from virions were present in approximately 1:1 molar ratios, the segments of the isolated cRNA were present in unequal amounts, with the segments M2 and S2 predominating, suggesting that different amounts of the cRNA segments were synthesized in the infected cell. The predominant cRNA segments, M2 and S2, and also the S1 segment, were active as mRNA's in wheat germ extracts. The M2 cRNA was the mRNA for the nucleocapsid protein; S1 for the membrane protein; and S2 for the nonstructural protein NS1.
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- Atkins J. F., Lewis J. B., Anderson C. W., Gesteland R. F. Enhanced differential synthesis of proteins in a mammalian cell-free system by addition of polyamines. J Biol Chem. 1975 Jul 25;250(14):5688–5695. [PubMed] [Google Scholar]
- Bean W. J., Jr, Simpson R. W. Transcriptase activity and genome composition of defective influenza virus. J Virol. 1976 Apr;18(1):365–369. doi: 10.1128/jvi.18.1.365-369.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Buchhagen D. L., Stutman O., Fleissner E. Chromatographic Separation and Antigenic Analysis of Proteins of the Oncornaviruses IV. Biochemical Typing of Murine Viral Proteins. J Virol. 1975 May;15(5):1148–1157. doi: 10.1128/jvi.15.5.1148-1157.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Content J. Cell-free translation of influenza virus mRNA. J Virol. 1976 May;18(2):604–618. doi: 10.1128/jvi.18.2.604-618.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Etkind P. R., Krug R. M. Influenza viral messenger RNA. Virology. 1974 Nov;62(1):38–45. doi: 10.1016/0042-6822(74)90301-8. [DOI] [PubMed] [Google Scholar]
- Etkind P. R., Krug R. M. Purification of influenza viral complementary RNA: its genetic content and activity in wheat germ cell-free extracts. J Virol. 1975 Dec;16(6):1464–1475. doi: 10.1128/jvi.16.6.1464-1475.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Franklin R. M. Purification and properties of the replicative intermediate of the RNA bacteriophage R17. Proc Natl Acad Sci U S A. 1966 Jun;55(6):1504–1511. doi: 10.1073/pnas.55.6.1504. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Glass S. E., McGeoch D., Barry R. D. Characterization of the mRNA of influenza virus. J Virol. 1975 Dec;16(6):1435–1443. doi: 10.1128/jvi.16.6.1435-1443.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Horst J., Content J., Mandeles S., Fraenkel-Conrat H., Duesberg P. Distinct oligonucleotide patterns of distinct influenza virus RNA's. J Mol Biol. 1972 Aug 21;69(2):209–215. doi: 10.1016/0022-2836(72)90226-4. [DOI] [PubMed] [Google Scholar]
- Inglis S. C., Carroll A. R., Lamb R. A., Mahy B. W. Polypeptides specified by the influenza virus genome I. Evidence for eight distinct gene products specified by fowl plague virus. Virology. 1976 Oct 15;74(2):489–503. doi: 10.1016/0042-6822(76)90355-x. [DOI] [PubMed] [Google Scholar]
- Klenk H. D., Rott R. Formation of influenza virus proteins. J Virol. 1973 Jun;11(6):823–831. doi: 10.1128/jvi.11.6.823-831.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Krug R. M. Cytoplasmic and nucleoplasmic viral RNPs in influenza virus-infected MDCK cells. Virology. 1972 Oct;50(1):103–113. doi: 10.1016/0042-6822(72)90350-9. [DOI] [PubMed] [Google Scholar]
- Krug R. M., Etkind P. R. Cytoplasmic and nuclear virus-specific proteins in influenza virus-infected MDCK cells. Virology. 1973 Nov;56(1):334–348. doi: 10.1016/0042-6822(73)90310-3. [DOI] [PubMed] [Google Scholar]
- Krug R. M. Influenza viral RNPs newly synthesized during the latent period of viral growth in MDCK cells. Virology. 1971 Apr;44(1):125–136. doi: 10.1016/0042-6822(71)90159-0. [DOI] [PubMed] [Google Scholar]
- Krug R. M., Morgan M. A., Shatkin A. J. Influenza viral mRNA contains internal N6-methyladenosine and 5'-terminal 7-methylguanosine in cap structures. J Virol. 1976 Oct;20(1):45–53. doi: 10.1128/jvi.20.1.45-53.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lamb R. A., Choppin P. W. Synthesis of influenza virus proteins in infected cells: translation of viral polypeptides, including three P polypeptides, from RNA produced by primary transcription. Virology. 1976 Oct 15;74(2):504–519. doi: 10.1016/0042-6822(76)90356-1. [DOI] [PubMed] [Google Scholar]
- Lazarowitz S. G., Compans R. W., Choppin P. W. Influenza virus structural and nonstructural proteins in infected cells and their plasma membranes. Virology. 1971 Dec;46(3):830–843. doi: 10.1016/0042-6822(71)90084-5. [DOI] [PubMed] [Google Scholar]
- Lazarowitz S. G., Goldberg A. R., Choppin P. W. Proteolytic cleavage by plasmin of the HA polypeptide of influenza virus: host cell activation of serum plasminogen. Virology. 1973 Nov;56(1):172–180. doi: 10.1016/0042-6822(73)90296-1. [DOI] [PubMed] [Google Scholar]
- McGeoch D., Fellner P., Newton C. Influenza virus genome consists of eight distinct RNA species. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3045–3049. doi: 10.1073/pnas.73.9.3045. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meier-Ewert H., Compans R. W. Time course of synthesis and assembly of influenza virus proteins. J Virol. 1974 Nov;14(5):1083–1091. doi: 10.1128/jvi.14.5.1083-1091.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Palese P., Schulman J. L. Mapping of the influenza virus genome: identification of the hemagglutinin and the neuraminidase genes. Proc Natl Acad Sci U S A. 1976 Jun;73(6):2142–2146. doi: 10.1073/pnas.73.6.2142. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Plotch S. J., Krug R. M. Influenza virion transcriptase: synthesis in vitro of large, polyadenylic acid-containing complementary RNA. J Virol. 1977 Jan;21(1):24–34. doi: 10.1128/jvi.21.1.24-34.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pons M. W. A reexamination of influenza single-and double-stranded RNAs by gel electrophoresis. Virology. 1976 Feb;69(2):789–792. doi: 10.1016/0042-6822(76)90508-0. [DOI] [PubMed] [Google Scholar]
- Richey M. B., Palese P. In vitro translation of influenza virus messenger RNAs. Virology. 1976 Jul 15;72(2):410–420. doi: 10.1016/0042-6822(76)90170-7. [DOI] [PubMed] [Google Scholar]
- Ritchey M. B., Palese P., Schulman J. L. Mapping of the influenza virus genome. III. Identification of genes coding for nucleoprotein, membrane protein, and nonstructural protein. J Virol. 1976 Oct;20(1):307–313. doi: 10.1128/jvi.20.1.307-313.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scholtissek C., Harms E., Rohde W., Orlich M., Rott R. Correlation between RNA fragments of fowl plague virus and their corresponding gene functions. Virology. 1976 Oct 15;74(2):332–344. doi: 10.1016/0042-6822(76)90340-8. [DOI] [PubMed] [Google Scholar]
- Schuerch A. R., Mitchell W. R., Joklik W. K. Isolation of intact individual species of single- and double-stranded RNA after fractionation by polyacrylamide gel electrophoresis. Anal Biochem. 1975 May 12;65(1-2):331–345. doi: 10.1016/0003-2697(75)90517-5. [DOI] [PubMed] [Google Scholar]
- Skehel J. J. Polypeptide synthesis in influenza virus-infected cells. Virology. 1972 Jul;49(1):23–36. doi: 10.1016/s0042-6822(72)80004-7. [DOI] [PubMed] [Google Scholar]
- Stavrianopoulos J. G., Chargaff E. Purification and properties of ribonuclease H of calf thymus. Proc Natl Acad Sci U S A. 1973 Jul;70(7):1959–1963. doi: 10.1073/pnas.70.7.1959. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stavrianopoulos J. G., Gambino-Giuffrida A., Chargaff E. Ribonuclease H of calf thymus: substrate specificity, activation, inhibition. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1087–1091. doi: 10.1073/pnas.73.4.1087. [DOI] [PMC free article] [PubMed] [Google Scholar]