Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1977 Dec;24(3):786–793. doi: 10.1128/jvi.24.3.786-793.1977

Two methyltransferase activities in the purified virions of vesicular stomatitis virus.

D Testa, A K Banerjee
PMCID: PMC516000  PMID: 201777

Abstract

In addition to an RNA-dependent RNA polymerase, purified vesicular stomatitis virus contains a methyltransferase activity which transfers the methyl group from the methyl donor, S-adenosyl-L-methionine, to two positions in the 5'-terminal capped structure of the nascent mRNA's synthesized in vitro as 7mG-(5)'ppp(5')Apm... In the present study it is shown that two distinct methyltransferase activities are discernible in the purified virus. The in vitro concentrations of the methyl donor specify the number and location of the methyl groups transferred to the capped 5'-termini of VSV mRNA's. Limited concentrations of the methyl donor result in a single methylation of the penultimate base in the 2'-hydroxyl position, that is, G(5')ppp(5')Apm..., whereas saturating concentrations of the methyl donor methylate the blocking guanosine residue at the 7-position, resulting in the dimethylated cap, 7mG(5')ppp(5')Apm... Pulse-chase experiments demonstrate that the monomethylated cap structure is the precursor substrate for the dimethylated cap. In this respect, vesicular stomatitis virus system is quite distinct from the vaccinia and reovirus systems. Virus purified from different host cells including hamster, mouse, and human contain both methyltransferase activities. The mRNA's containing monomethylated capped structures are poor templates for protein synthesis in vitro.

Full text

PDF
786

Selected References

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

  1. Abraham G., Banerjee A. K. The nature of the RNA products synthesized in vitro by subviral components of visicular stomatitis virus. Virology. 1976 May;71(1):230–241. doi: 10.1016/0042-6822(76)90108-2. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Baltimore D., Huang A. S., Stampfer M. Ribonucleic acid synthesis of vesicular stomatitis virus, II. An RNA polymerase in the virion. Proc Natl Acad Sci U S A. 1970 Jun;66(2):572–576. doi: 10.1073/pnas.66.2.572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Banerjee A. K., Moyer S. A., Rhodes D. P. Studies on the in vitro adenylation of RNA by vesicular stomatitis virus. Virology. 1974 Oct;61(2):547–558. doi: 10.1016/0042-6822(74)90289-x. [DOI] [PubMed] [Google Scholar]
  5. Banerjee A. K., Rhodes D. P. In vitro synthesis of RNA that contains polyadenylate by virion-associated RNA polymerase of vesicular stomatitis virus. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3566–3570. doi: 10.1073/pnas.70.12.3566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Both G. W., Banerjee A. K., Shatkin A. J. Methylation-dependent translation of viral messenger RNAs in vitro. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1189–1193. doi: 10.1073/pnas.72.3.1189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Colonno R. J., Banerjee A. K. A unique RNA species involved in initiation of vesicular stomatitis virus RNA transcription in vitro. Cell. 1976 Jun;8(2):197–204. doi: 10.1016/0092-8674(76)90003-9. [DOI] [PubMed] [Google Scholar]
  10. Colonno R. J., Banerjee A. K. Mapping and initiation studies on the leader RNA of vesicular stomatitis virus. Virology. 1977 Mar;77(1):260–268. doi: 10.1016/0042-6822(77)90423-8. [DOI] [PubMed] [Google Scholar]
  11. Colonno R. J., Stone H. O. Newcastle disease virus mRNA lacks 2'-O-methylated nucleotides. Nature. 1976 Jun 17;261(5561):611–614. doi: 10.1038/261611a0. [DOI] [PubMed] [Google Scholar]
  12. Emerson S. U., Yu Y. Both NS and L proteins are required for in vitro RNA synthesis by vesicular stomatitis virus. J Virol. 1975 Jun;15(6):1348–1356. doi: 10.1128/jvi.15.6.1348-1356.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ensinger M. J., Martin S. A., Paoletti E., Moss B. Modification of the 5'-terminus of mRNA by soluble guanylyl and methyl transferases from vaccinia virus. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2525–2529. doi: 10.1073/pnas.72.7.2525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. 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]
  17. Hickey E. D., Weber L. A., Baglioni C. Inhibition of initiation of protein synthesis by 7-methylguanosine-5'-monophosphate. Proc Natl Acad Sci U S A. 1976 Jan;73(1):19–23. doi: 10.1073/pnas.73.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Imblum R. L., Wagner R. R. Inhibition of viral transcriptase by immunoglobulin directed against the nucleocapsid NS protein of vesicular stomatitis virus. J Virol. 1975 Jun;15(6):1357–1366. doi: 10.1128/jvi.15.6.1357-1366.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Martin S. A., Moss B. Modification of RNA by mRNA guanylyltransferase and mRNA (guanine-7-)methyltransferase from vaccinia virions. J Biol Chem. 1975 Dec 25;250(24):9330–9335. [PubMed] [Google Scholar]
  20. Martin S. A., Paoletti E., Moss B. Purification of mRNA guanylyltransferase and mRNA (guanine-7-) methyltransferase from vaccinia virions. J Biol Chem. 1975 Dec 25;250(24):9322–9329. [PubMed] [Google Scholar]
  21. Moss B., Gershowitz A., Wei C. M., Boone R. Formation of the guanylylated and methylated 5'-terminus of vaccinia virus mRNA. Virology. 1976 Jul 15;72(2):341–351. doi: 10.1016/0042-6822(76)90163-x. [DOI] [PubMed] [Google Scholar]
  22. Moyer S. A., Banerjee A. K. In vivo methylation of vesicular stomatitis virus and its host-cell messenger RNA species. Virology. 1976 Apr;70(2):339–351. doi: 10.1016/0042-6822(76)90276-2. [DOI] [PubMed] [Google Scholar]
  23. Muthukrishnan S., Filipowicz W., Sierra J. M., Both G. W., Shatkin A. J., Ochoa S. mRNA methylation and protein synthesis in extracts from embryos of brine shrimp, Artemia salina. J Biol Chem. 1975 Dec 25;250(24):9336–9341. [PubMed] [Google Scholar]
  24. Muthukrishnan S., Morgan M., Banerjee A. K., Shatkin A. J. Influence of 5'-terminal m7G and 2'--O-methylated residues on messenger ribonucleic acid binding to ribosomes. Biochemistry. 1976 Dec 28;15(26):5761–5768. doi: 10.1021/bi00671a012. [DOI] [PubMed] [Google Scholar]
  25. Rhodes D. P., Banerjee A. K. 5'-terminal sequence of vesicular stomatitis virus mRNA's synthesized in vitro. J Virol. 1975 Jan;17(1):33–42. doi: 10.1128/jvi.17.1.33-42.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rhodes D. P., Reddy D. V., MacLeod R., Black L. M., Banerjee A. K. In vitro synthesis of RNA containing 5'-terminal structure 7nG(5')ppp(5')Apm... by purified wound tumor virus. Virology. 1977 Feb;76(2):554–559. doi: 10.1016/0042-6822(77)90237-9. [DOI] [PubMed] [Google Scholar]
  27. 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]

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

RESOURCES