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. 1982 Jul;43(1):234–240. doi: 10.1128/jvi.43.1.234-240.1982

Capping of Viral RNA in Cultured Spodoptera frugiperda Cells Infected with Autographa californica Nuclear Polyhedrosis Virus

Qin Jun-Chuan 1,, Robert F Weaver 1
PMCID: PMC256114  PMID: 16789227

Abstract

Viral RNA from fall armyworm (Spodoptera frugiperda) cells infected with Autographa californica nuclear polyhedrosis virus contains cap structures. Most of the cap labeled in vivo with [3H]methionine or 32Pi cochromatographed on DEAE-cellulose with the −5 charge marker; a minor component appeared at −4 net charge. The former is probably a cap 1 structure (m7GpppXmpYp), and the latter is probably a cap 0 (m7GpppXp). On the basis of relative labeling of the two caps with [3H]adenosine and [3H]guanosine, we concluded that each cap contained at least one adenosine residue in addition to guanosine and, therefore, that cap 0 contained m7GpppAp. Cleavage of [3H]methionine-labeled viral RNA with tobacco acid pyrophosphatase released a labeled component that cochromatographed on polyethyleneimine-cellulose with m7Gp, confirming the m7GpppX linkage in the cap. These results were also seen with host polyadenylated RNA. The caps appeared in nearly equal abundance in viral polyadenylated and non-polyadenylated RNAs. The ratio of 32Pi incorporated into the cap to that incorporated into mononucleotides suggested average lengths for the polyadenylated and non-polyadenylated RNAs of 1,800 and 1,200 nucleotides, respectively.

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Selected References

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  1. Adams J. M., Cory S. Modified nucleosides and bizarre 5'-termini in mouse myeloma mRNA. Nature. 1975 May 1;255(5503):28–33. doi: 10.1038/255028a0. [DOI] [PubMed] [Google Scholar]
  2. Colonno R. J., Stone H. O. Methylation of messenger RNA of Newcastle disease virus in vitro by a virion-associated enzyme. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2611–2615. doi: 10.1073/pnas.72.7.2611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cory S., Adams J. M. The modified 5'-terminal sequences in messenger RNA of mouse myeloma cells. J Mol Biol. 1975 Dec 25;99(4):519–547. doi: 10.1016/s0022-2836(75)80170-7. [DOI] [PubMed] [Google Scholar]
  4. Dasgupta R., Harada F., Kaesberg P. Blocked 5' termini in brome mosaic virus RNA. J Virol. 1976 Apr;18(1):260–267. doi: 10.1128/jvi.18.1.260-267.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dottin R. P., Weiner A. M., Lodish F. 5' terminal nucleotide sequences of the messenger RNA's of Dictyostelium discoideum. Cell. 1976 Jun;8(2):233–244. doi: 10.1016/0092-8674(76)90007-6. [DOI] [PubMed] [Google Scholar]
  6. Dubin D. T., Stollar V. Methylation of Sindbis virus "26S" messenger RNA. Biochem Biophys Res Commun. 1975 Oct 27;66(4):1373–1379. doi: 10.1016/0006-291x(75)90511-2. [DOI] [PubMed] [Google Scholar]
  7. Furuichi Y., Miura K. A blocked structure at the 5' terminus of mRNA from cytoplasmic polyhedrosis virus. Nature. 1975 Jan 31;253(5490):374–375. doi: 10.1038/253374a0. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Goodwin R. H. Insect cell culture: improved media and methods for initiating attached cell lines from the Lepidoptera. In Vitro. 1975 Nov-Dec;11(6):369–378. doi: 10.1007/BF02616373. [DOI] [PubMed] [Google Scholar]
  10. Grula M. A., Buller P. L., Weaver R. F. alpha-Amanitin-Resistant Viral RNA Synthesis in Nuclei Isolated from Nuclear Polyhedrosis Virus-Infected Heliothis zea Larvae and Spodoptera frugiperda Cells. J Virol. 1981 Jun;38(3):916–921. doi: 10.1128/jvi.38.3.916-921.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Milcarek C. HeLa cell cytoplasmic mRNA contains three classes of sequences: predominantly poly(A)-free, predominantly poly(A)-containing and bimorphic. Eur J Biochem. 1979 Dec 17;102(2):467–476. doi: 10.1111/j.1432-1033.1979.tb04261.x. [DOI] [PubMed] [Google Scholar]
  12. Miller L. K., Dawes K. P. Physical Map of the DNA Genome of Autographa californica Nuclear Polyhedrosis Virus. J Virol. 1979 Mar;29(3):1044–1055. doi: 10.1128/jvi.29.3.1044-1055.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Noyes B. E., Stark G. R. Nucleic acid hybridization using DNA covalently coupled to cellulose. Cell. 1975 Jul;5(3):301–310. doi: 10.1016/0092-8674(75)90105-1. [DOI] [PubMed] [Google Scholar]
  14. Ouellette A. J. Purification by benzoylated cellulose chromatography of translatable messenger ribonucleic acid lacking polyadenylate. J Biol Chem. 1980 Apr 10;255(7):2740–2746. [PubMed] [Google Scholar]
  15. Palmiter R. D. Magnesium precipitation of ribonucleoprotein complexes. Expedient techniques for the isolation of undergraded polysomes and messenger ribonucleic acid. Biochemistry. 1974 Aug 13;13(17):3606–3615. doi: 10.1021/bi00714a032. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Shatkin A. J. Capping of eucaryotic mRNAs. Cell. 1976 Dec;9(4 Pt 2):645–653. doi: 10.1016/0092-8674(76)90128-8. [DOI] [PubMed] [Google Scholar]
  18. Sripati C. E., Groner Y., Warner J. R. Methylated, blocked 5' termini of yeast mRNA. J Biol Chem. 1976 May 25;251(10):2898–2904. [PubMed] [Google Scholar]
  19. 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]

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