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. 1982 Mar;41(3):1105–1108. doi: 10.1128/jvi.41.3.1105-1108.1982

mRNA 5'-cap binding activity in purified influenza virus detected by simple, rapid assay.

H Kroath, A J Shatkin
PMCID: PMC256851  PMID: 7097854

Abstract

Reovirus mRNA 5'-terminal caps were 3'-radiolabeled with pCp and as affinity probes for proteins with cap binding activity. A rapid, simple, and sensitive blot assay was devised that could detect cellular cap binding protein in a complex polypeptide mixture. By using this method, cap binding activity was found in detergent-treated influenza virus but not in reovirus or vaccinia virus. Preincubation of capped reovirus mRNA with purified cellular cap binding protein reduced its primer effect on influenza transcriptase, whereas priming by ApG was not affected. The results indicate that influenza transcriptase complexes include cap-recognizing proteins that are involved in the formation of chimeric mRNAs.

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Selected References

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  1. Bergmann J. E., Trachsel H., Sonenberg N., Shatkin A. J., Lodish H. F. Characterization of rabbit reticulocyte factor(s) that stimulates the translation of mRNAs lacking 5'-terminal 7-methylguanosine. J Biol Chem. 1979 Mar 10;254(5):1440–1443. [PubMed] [Google Scholar]
  2. Both G. W., Furuichi Y., Muthukrishnan S., Shatkin A. J. Effect of 5'-terminal structure and base composition on polyribonucleotide binding to ribosomes. J Mol Biol. 1976 Jul 5;104(3):637–658. doi: 10.1016/0022-2836(76)90126-1. [DOI] [PubMed] [Google Scholar]
  3. England T. E., Uhlenbeck O. C. Enzymatic oligoribonucleotide synthesis with T4 RNA ligase. Biochemistry. 1978 May 30;17(11):2069–2076. doi: 10.1021/bi00604a008. [DOI] [PubMed] [Google Scholar]
  4. Filipowicz W., Furuichi Y., Sierra J. M., Muthukrishnan S., Shatkin A. J., Ochoa S. A protein binding the methylated 5'-terminal sequence, m7GpppN, of eukaryotic messenger RNA. Proc Natl Acad Sci U S A. 1976 May;73(5):1559–1563. doi: 10.1073/pnas.73.5.1559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Furuichi Y., Muthukrishnan S., Tomasz J., Shatkin A. J. Mechanism of formation of reovirus mRNA 5'-terminal blocked and methylated sequence, m7GpppGmpC. J Biol Chem. 1976 Aug 25;251(16):5043–5053. [PubMed] [Google Scholar]
  6. 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]
  7. JOKLIK W. K. The preparation and characteristics of highly purified radioactively labelled poxvirus. Biochim Biophys Acta. 1962 Aug 20;61:290–301. doi: 10.1016/0926-6550(62)90091-9. [DOI] [PubMed] [Google Scholar]
  8. Krug R. M., Broni B. A., LaFiandra A. J., Morgan M. A., Shatkin A. J. Priming and inhibitory activities of RNAs for the influenza viral transcriptase do not require base pairing with the virion template RNA. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5874–5878. doi: 10.1073/pnas.77.10.5874. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Krug R. M. Priming of influenza viral RNA transcription by capped heterologous RNAs. Curr Top Microbiol Immunol. 1981;93:125–149. doi: 10.1007/978-3-642-68123-3_6. [DOI] [PubMed] [Google Scholar]
  11. Lerner M. R., Boyle J. A., Mount S. M., Wolin S. L., Steitz J. A. Are snRNPs involved in splicing? Nature. 1980 Jan 10;283(5743):220–224. doi: 10.1038/283220a0. [DOI] [PubMed] [Google Scholar]
  12. Martin S. A., Moss B. mRNA guanylyltransferase and mRNA (guanine-7-)-methyltransferase from vaccinia virions. Donor and acceptor substrate specificites. J Biol Chem. 1976 Dec 10;251(23):7313–7321. [PubMed] [Google Scholar]
  13. 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]
  14. Plotch S. J., Bouloy M., Ulmanen I., Krug R. M. A unique cap(m7GpppXm)-dependent influenza virion endonuclease cleaves capped RNAs to generate the primers that initiate viral RNA transcription. Cell. 1981 Mar;23(3):847–858. doi: 10.1016/0092-8674(81)90449-9. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Shatkin A. J., LaFiandra A. J. Transcription by infectious subviral particles of reovirus. J Virol. 1972 Oct;10(4):698–706. doi: 10.1128/jvi.10.4.698-706.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Smith J. C., Raper R. H., Bell L. D., Stebbing N., McGeoch D. Inhibition of influenza virion transcriptases by polynucleotides. Virology. 1980 May;103(1):245–249. doi: 10.1016/0042-6822(80)90144-0. [DOI] [PubMed] [Google Scholar]
  18. Sonenberg N., Rupprecht K. M., Hecht S. M., Shatkin A. J. Eukaryotic mRNA cap binding protein: purification by affinity chromatography on sepharose-coupled m7GDP. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4345–4349. doi: 10.1073/pnas.76.9.4345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sonenberg N., Shatkin A. J. Reovirus mRNA can be covalently crosslinked via the 5' cap to proteins in initiation complexes. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4288–4292. doi: 10.1073/pnas.74.10.4288. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tahara S. M., Morgan M. A., Shatkin A. J. Two forms of purified m7G-cap binding protein with different effects on capped mRNA translation in extracts of uninfected and poliovirus-infected HeLa cells. J Biol Chem. 1981 Aug 10;256(15):7691–7694. [PubMed] [Google Scholar]

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