Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1976 Oct;20(1):45–53. doi: 10.1128/jvi.20.1.45-53.1976

Influenza viral mRNA contains internal N6-methyladenosine and 5'-terminal 7-methylguanosine in cap structures.

R M Krug, M A Morgan, A J Shatkin
PMCID: PMC354964  PMID: 1086370

Abstract

Influenza viral complementary RNA (cRNA), i.e., viral mRNA was radioactive when purified from the cytoplasmic fraction of cordycepin-treated canine kidney cells that were incubated with [methyl-3H]methionine during infection. Approximately 55 to 60% of the methyl-3H radioactivity was in internal N6-methyladenosine, a feature distinguishing this mRNA from those viral mRNA's that are known to be synthesized in the cytoplasm. The remaining methyl-3H radioactivity was in 5'-terminal cap structures that consisted of 7-methylguanosine in pyrophosphate linkage to 2'-o-methyladenosine, N6, 2'-O-dimethyladenosine, or 2'-O-methylguanosine. Methylated adenosine was the predominant penultimate nucleoside in caps, suggesting that cRNA synthesis in infected cells initiates preferentially with adenosine at the 5' end. In contrast to cRNA, influenza virion RNA segments extracted from purified virus contained mainly 5'-terminal ppA and no detectable cap structures.

Full text

PDF
45

Selected References

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

  1. Abraham G., Rhodes D. P., Banerjee A. K. The 5' terminal structure of the methylated mRNA synthesized in vitro by vesicular stomatitis virus. Cell. 1975 May;5(1):51–58. doi: 10.1016/0092-8674(75)90091-4. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Bishop D. H., Obijeski J. F., Simpson R. W. Transcription of the influenza ribonucleic acid genome by a virion polymerase. II. Nature of the in vitro polymerase product. J Virol. 1971 Jul;8(1):74–80. doi: 10.1128/jvi.8.1.74-80.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cox N. J., Barry R. D. Hybridization studies of the relationship between influenza virus RNA and cellular DNA. Virology. 1976 Jan;69(1):304–313. doi: 10.1016/0042-6822(76)90217-8. [DOI] [PubMed] [Google Scholar]
  5. Desrosiers R. C., Friderici K. H., Rottman F. M. Characterization of Novikoff hepatoma mRNA methylation and heterogeneity in the methylated 5' terminus. Biochemistry. 1975 Oct 7;14(20):4367–4374. doi: 10.1021/bi00691a004. [DOI] [PubMed] [Google Scholar]
  6. Dimmock N. J. New virus-specific antigens in cells infected with influenza virus. Virology. 1969 Oct;39(2):224–234. doi: 10.1016/0042-6822(69)90042-7. [DOI] [PubMed] [Google Scholar]
  7. Erikson R. L., Gordon J. A. Replication of bacteriophage RNA: purification of the replicative intermediate by agarose column chromatography-1. Biochem Biophys Res Commun. 1966 May 25;23(4):422–428. doi: 10.1016/0006-291x(66)90744-3. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Follett E. A., Pringle C. R., Wunner W. H., Skehel J. J. Virus replication in enucleate cells: vesicular stomatitis virus and influenza virus. J Virol. 1974 Feb;13(2):394–399. doi: 10.1128/jvi.13.2.394-399.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Furuichi Y., Morgan M., Muthukrishnan S., Shatkin A. J. Reovirus messenger RNA contains a methylated, blocked 5'-terminal structure: m-7G(5')ppp(5')G-MpCp-. Proc Natl Acad Sci U S A. 1975 Jan;72(1):362–366. doi: 10.1073/pnas.72.1.362. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Furuichi Y., Morgan M., Shatkin A. J., Jelinek W., Salditt-Georgieff M., Darnell J. E. Methylated, blocked 5 termini in HeLa cell mRNA. Proc Natl Acad Sci U S A. 1975 May;72(5):1904–1908. doi: 10.1073/pnas.72.5.1904. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ghendon Y., Blagoveshienskaya O. Polyadenylate sequences of fowl plague virus complementary RNA (cRNA) synthesized in vivo and in vitro. Virology. 1975 Dec;68(2):330–337. doi: 10.1016/0042-6822(75)90276-7. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Keith J., Gleason M., Fraenkel-Conrat H. Characterization of the end groups of RNA of Rous sarcoma virus. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4371–4375. doi: 10.1073/pnas.71.11.4371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kelly D. C., Avery R. J., Dimmock N. J. Failure of an influenza virus to initiate infection in enucleate BHK cells. J Virol. 1974 Jun;13(6):1155–1161. doi: 10.1128/jvi.13.6.1155-1161.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. Krug R. M., Soeiro R. Studies on the intranuclear localization of influenza virus-specific proteins. Virology. 1975 Apr;64(2):378–387. doi: 10.1016/0042-6822(75)90114-2. [DOI] [PubMed] [Google Scholar]
  20. Lavi S., Shatkin A. J. Methylated simian virus 40-specific RNA from nuclei and cytoplasm of infected BSC-1 cells. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2012–2016. doi: 10.1073/pnas.72.6.2012. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Lewandowski L. J., Content J., Leppla S. H. Characterization of the subunit structure of the ribonucleic acid genome of influenza virus. J Virol. 1971 Nov;8(5):701–707. doi: 10.1128/jvi.8.5.701-707.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. McGeoch D., Kitron N. Influenza virion RNA-dependent RNA polymerase: stimulation by guanosine and related compounds. J Virol. 1975 Apr;15(4):686–695. doi: 10.1128/jvi.15.4.686-695.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Moss B., Koczot F. Sequence of methylated nucleotides at the 5'-terminus of adenovirus-specific RNA. J Virol. 1976 Feb;17(2):385–392. doi: 10.1128/jvi.17.2.385-392.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Perry R. P., Kelley D. E., Friderici K., Rottman F. The methylated constituents of L cell messenger RNA: evidence for an unusual cluster at the 5' terminus. Cell. 1975 Apr;4(4):387–394. doi: 10.1016/0092-8674(75)90159-2. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Pons M. W. The inhibition of influenza virus RNA synthesis by actinomycin D and cycloheximide. Virology. 1973 Jan;51(1):120–128. doi: 10.1016/0042-6822(73)90372-3. [DOI] [PubMed] [Google Scholar]
  28. Rose J. K. Heterogneeous 5'-terminal structures occur on vesicular stomatitis virus mRNAs. J Biol Chem. 1975 Oct 25;250(20):8098–8104. [PubMed] [Google Scholar]
  29. Scholtissek C., Rott R. Synthesis in vivo of influenza virus plus and minus strand RNA and its preferential inhibition by antibiotics. Virology. 1970 Apr;40(4):989–996. doi: 10.1016/0042-6822(70)90145-5. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Taylor J. M., Hampson A. W., White D. O. The polypeptides of influenza virus. 1. Cytoplasmic synthesis and nuclear accumulation. Virology. 1969 Nov;39(3):419–425. doi: 10.1016/0042-6822(69)90090-7. [DOI] [PubMed] [Google Scholar]
  32. Wei C. M., Gershowitz A., Moss B. 5'-Terminal and internal methylated nucleotide sequences in HeLa cell mRNA. Biochemistry. 1976 Jan 27;15(2):397–401. doi: 10.1021/bi00647a024. [DOI] [PubMed] [Google Scholar]
  33. Wei C. M., Gershowitz A., Moss B. Methylated nucleotides block 5' terminus of HeLa cell messenger RNA. Cell. 1975 Apr;4(4):379–386. doi: 10.1016/0092-8674(75)90158-0. [DOI] [PubMed] [Google Scholar]
  34. Wei C. M., Moss B. Methylated nucleotides block 5'-terminus of vaccinia virus messenger RNA. Proc Natl Acad Sci U S A. 1975 Jan;72(1):318–322. doi: 10.1073/pnas.72.1.318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Young R. J., Content J. 5'-terminus of influenza virus RNA. Nat New Biol. 1971 Mar 31;230(13):140–142. doi: 10.1038/newbio230140a0. [DOI] [PubMed] [Google Scholar]

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

RESOURCES