Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1975 Mar;72(3):1189–1193. doi: 10.1073/pnas.72.3.1189

Methylation-dependent translation of viral messenger RNAs in vitro.

G W Both, A K Banerjee, A J Shatkin
PMCID: PMC432492  PMID: 165487

Abstract

Methylated reovirus and vesicular stomatitis virus mRNAs, synthesized in vitro in the presence of S-adenosylmethionine by the virion-associated polymerases (RNA nucleotidyltransferases, EC 2.7.7.6), stimulate protein synthesis by wehat germ extracts to a greater extent than unmethylated mRNAs. Addition of S-adenosylmethionine to a cell-free extract programmed with unmethylated mRNA stimulates protein synthesis and results in methylation of the mRNA. An inhibitor of mRNA methylation. S-adenosylhomocysteine, blocks translation of unmethylated, but not of methylated, mRNAs. Aurintricarboxylic acid, which inhibits polypepetide chain initiation, also prevents mRNA methylation by wheat germ extracts. In contrast, sparsomycin, which inhibits polypeptide chain elongation, does not reduce mRNA methylation. The results indicate that methylation of viral mRNA is required for translation in vitro and suggest that mRNA methylation occurs at the initiation step of protein synthesis.

Full text

PDF
1189

Selected References

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

  1. Banerjee A. K., Shatkin A. J. Transcription in vitro by reovirus-associated ribonucleic acid-dependent polymerase. J Virol. 1970 Jul;6(1):1–11. doi: 10.1128/jvi.6.1.1-11.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Both G. W., Lavi S., Shatkin A. J. Synthesis of all the gene products of the reovirus genome in vivo and in vitro. Cell. 1975 Feb;4(2):173–180. doi: 10.1016/0092-8674(75)90124-5. [DOI] [PubMed] [Google Scholar]
  3. Both G. W., Moyer S. A., Banerjee A. K. Translation and identification of the mRNA species synthesized in vitro by the virion-associated RNA polymerase of vesicular stomatitis virus. Proc Natl Acad Sci U S A. 1975 Jan;72(1):274–278. doi: 10.1073/pnas.72.1.274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Both G. W., Moyer S. A., Banerjee A. K. Translation and identification of the viral mRNA species isolated from subcellular fractions of vesicular stomatitis virus-infected cells. J Virol. 1975 Apr;15(4):1012–1019. doi: 10.1128/jvi.15.4.1012-1019.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Crippa M., Davidson E. H., Mirsky A. E. Persistence in early amphibian embryos of informational RNA's from the lampbrush chromosome stage of oögenesis. Proc Natl Acad Sci U S A. 1967 Apr;57(4):885–892. doi: 10.1073/pnas.57.4.885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Furuichi Y. "Methylation-coupled" transcription by virus-associated transcriptase of cytoplasmic polyhedrosis virus containing double-stranded RNA. Nucleic Acids Res. 1974 Jun;1(6):809–822. doi: 10.1093/nar/1.6.809. [DOI] [PMC free article] [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. Furuichi Y., Muthukrishnan S., Shatkin A. J. 5'-Terminal m-7G(5')ppp(5')G-m-p in vivo: identification in reovirus genome RNA. Proc Natl Acad Sci U S A. 1975 Feb;72(2):742–745. doi: 10.1073/pnas.72.2.742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Haselkorn R., Rothman-Denes L. B. Protein synthesis. Annu Rev Biochem. 1973;42:397–438. doi: 10.1146/annurev.bi.42.070173.002145. [DOI] [PubMed] [Google Scholar]
  11. Leong J. A., Garapin A. C., Jackson N., Fanshier L., Levinson W., Bishop J. M. Virus-specific ribonucleic acid in cells producing rous sarcoma virus: detection and characterization. J Virol. 1972 Jun;9(6):891–902. doi: 10.1128/jvi.9.6.891-902.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Levine L., Gjika H. Radioimmunoassays for N2-dimethylguanosine. 7-methylguanosine, and pseudouridine. Arch Biochem Biophys. 1974 Oct;164(2):583–589. doi: 10.1016/0003-9861(74)90070-8. [DOI] [PubMed] [Google Scholar]
  13. Marcus A., Bewley J. D., Weeks D. P. Aurintricarboxylic acid and initiation factors of wheat embryo. Science. 1970 Mar 27;167(3926):1735–1736. doi: 10.1126/science.167.3926.1735. [DOI] [PubMed] [Google Scholar]
  14. Moyer S. A., Banerjee A. K. Messenger RNA species synthesized in vitro by the virion-associated RNA polymerase of vesicular stomatitis virus. Cell. 1975 Jan;4(1):37–43. doi: 10.1016/0092-8674(75)90131-2. [DOI] [PubMed] [Google Scholar]
  15. Reddy R., Ro-Choi T. S., Henning D., Busch H. Primary sequence of U-1 nuclear ribonucleic acid of Novikoff hepatoma ascites cells. J Biol Chem. 1974 Oct 25;249(20):6486–6494. [PubMed] [Google Scholar]
  16. Rhodes D. P., Moyer S. A., Banerjee A. K. In vitro synthesis of methylated messenger RNA by the virion-associated RNA polymerase of vesicular stomatitis virus. Cell. 1974 Dec;3(4):327–333. doi: 10.1016/0092-8674(74)90046-4. [DOI] [PubMed] [Google Scholar]
  17. Rottman F., Shatkin A. J., Perry R. P. Sequences containing methylated nucleotides at the 5' termini of messenger RNAs: possible implications for processing. Cell. 1974 Nov;3(3):197–199. doi: 10.1016/0092-8674(74)90131-7. [DOI] [PubMed] [Google Scholar]
  18. Shatkin A. J. Animal RNA viruses: genome structure and function. Annu Rev Biochem. 1974;43(0):643–665. doi: 10.1146/annurev.bi.43.070174.003235. [DOI] [PubMed] [Google Scholar]
  19. Shatkin A. J. Methylated messenger RNA synthesis in vitro by purified reovirus. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3204–3207. doi: 10.1073/pnas.71.8.3204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Silber R., Malathi V. G., Schulman L. H., Hurwitz J., Duesberg P. H. Studies of the Rous sarcoma virus RNA: characterization of the 5'-terminus. Biochem Biophys Res Commun. 1973 Jan 23;50(2):467–472. doi: 10.1016/0006-291x(73)90863-2. [DOI] [PubMed] [Google Scholar]
  21. Slater D. W., Spiegelman S. An estimation of genetic messages in the unfertilized echinoid egg. Proc Natl Acad Sci U S A. 1966 Jul;56(1):164–170. doi: 10.1073/pnas.56.1.164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Smith A. E., Wigle D. T. A rapid assay for the initiation of protein synthesis in extracts of animal cells. Eur J Biochem. 1973 Jun 15;35(3):566–573. doi: 10.1111/j.1432-1033.1973.tb02874.x. [DOI] [PubMed] [Google Scholar]
  23. Urushibara T., Furuichi Y., Nishimura C., Miura K. A modified structure at the 5'-terminus of mRNA of vaccinia virus. FEBS Lett. 1975 Jan 1;49(3):385–389. doi: 10.1016/0014-5793(75)80791-5. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Wei C. M., Moss B. Methylation of newly synthesized viral messenger RNA by an enzyme in vaccinia virus. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3014–3018. doi: 10.1073/pnas.71.8.3014. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES