Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1979 Jun 25;6(8):2879–2899. doi: 10.1093/nar/6.8.2879

Identification and mapping of N6-methyladenosine containing sequences in simian virus 40 RNA.

D Canaani, C Kahana, S Lavi, Y Groner
PMCID: PMC327900  PMID: 223130

Abstract

Late SV40 16S and 19S mRNAs were found to contain an average of three m6A residues per mRNA molecule. The methylated residues of both the viral and cellular mRNAs occur in two sequences; Gpm6ApC and (Ap)nm6ApC, where n = 1-4. More than 60% of the m6A residues in SV40 16S and 19S mRNAs occur in Gpm6ApC even though there are twice as many (A)nAC than GAC sequences in these messengers. The m6A containing oligonucleotides of late SV40 MRNAs were localized in the viral messengers. In the 16S mRNA two m6A oligonucleotides were located at the 5' coding region between 0.95--0.0 map units. The third m6A residue was mapped between 0.0--0.14 map units in the translated portion of this mRNA. The overall pattern of internal methylation in the 19S mRNA is similar. However, some differences between 16S and 19S mRNAs were observed in both the content and location of the longer (Ap)n m6AC nucleotides. These results provide the first example of precise localization of internal methylation sequences in mRNA species with defined coding specificity. It implies that a) location of m6A residues is not random but specific to a particular region of the RNA, b) apart from sequence specificity other structural features of the mRNA may influence internal methylation and c) m6A residues are present in coding regions of SV40 mRNAs.

Full text

PDF
2879

Images in this article

2892Image
on p.2892

Selected References

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

  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. Aloni Y. Biogenesis and characterization of SV40 and polyoma RNAs in productively infected cells. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):165–178. doi: 10.1101/sqb.1974.039.01.023. [DOI] [PubMed] [Google Scholar]
  3. Aloni Y., Dhar R., Laub O., Horowitz M., Khoury G. Novel mechanism for RNA maturation: the leader sequences of simian virus 40 mRNA are not transcribed adjacent to the coding sequences. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3686–3690. doi: 10.1073/pnas.74.9.3686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Aloni Y. Poly A and symmetrical transcription of SV40 DNA. Nat New Biol. 1973 May 2;243(122):2–6. [PubMed] [Google Scholar]
  5. Beemon K., Keith J. Localization of N6-methyladenosine in the Rous sarcoma virus genome. J Mol Biol. 1977 Jun 15;113(1):165–179. doi: 10.1016/0022-2836(77)90047-x. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Dhar R., Subramanian K. N., Pan J., Weissman S. M. Structure of a large segment of the genome of simian virus 40 that does not encode known proteins. Proc Natl Acad Sci U S A. 1977 Mar;74(3):827–831. doi: 10.1073/pnas.74.3.827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dimock K., Stoltzfus C. M. Sequence specificity of internal methylation in B77 avian sarcoma virus RNA subunits. Biochemistry. 1977 Feb 8;16(3):471–478. doi: 10.1021/bi00622a021. [DOI] [PubMed] [Google Scholar]
  9. Engel J. D., von Hippel P. H. Effects of methylation on the stability of nucleic acid conformations: studies at the monomer level. Biochemistry. 1974 Sep 24;13(20):4143–4158. doi: 10.1021/bi00717a013. [DOI] [PubMed] [Google Scholar]
  10. Fiers W., Contreras R., Haegemann G., Rogiers R., Van de Voorde A., Van Heuverswyn H., Van Herreweghe J., Volckaert G., Ysebaert M. Complete nucleotide sequence of SV40 DNA. Nature. 1978 May 11;273(5658):113–120. doi: 10.1038/273113a0. [DOI] [PubMed] [Google Scholar]
  11. Furuichi Y., Shatkin A. J., Stavnezer E., Bishop J. M. Blocked, methylated 5'-terminal sequence in avian sarcoma virus RNA. Nature. 1975 Oct 16;257(5527):618–620. doi: 10.1038/257618a0. [DOI] [PubMed] [Google Scholar]
  12. GRIFFIN B. E., HASLAM W. J., REESE C. B. SYNTHESIS AND PROPERTIES OF SOME METHYLATED POLYADENYLIC ACIDS. J Mol Biol. 1964 Nov;10:353–356. doi: 10.1016/s0022-2836(64)80055-3. [DOI] [PubMed] [Google Scholar]
  13. Gelinas R. E., Roberts R. J. One predominant 5'-undecanucleotide in adenovirus 2 late messenger RNAs. Cell. 1977 Jul;11(3):533–544. doi: 10.1016/0092-8674(77)90071-x. [DOI] [PubMed] [Google Scholar]
  14. Ghosh P. K., Reddy V. B., Swinscoe J., Choudary P. V., Lebowitz P., Weissman S. M. The 5'-terminal leader sequence of late 16 S mRNA from cells infected with simian virus 40. J Biol Chem. 1978 May 25;253(10):3643–3647. [PubMed] [Google Scholar]
  15. Gilboa E., Prives C. L., Aviv H. Purification of SV-40 messenger RNA by hybridization to SV-40 DNA covalently bound to Sepharose. Biochemistry. 1975 Sep 23;14(19):4215–4220. doi: 10.1021/bi00690a010. [DOI] [PubMed] [Google Scholar]
  16. Gilham P. T., Rosenberg M. The isolation of 3'-terminal polynucleotides from RNA molecules. Biochim Biophys Acta. 1971 Aug 26;246(2):337–340. doi: 10.1016/0005-2787(71)90143-2. [DOI] [PubMed] [Google Scholar]
  17. Groner Y., Carmi P., Aloni Y. Capping structures of simian virus 40 19S and 16S mRNAs. Nucleic Acids Res. 1977 Nov;4(11):3959–3968. doi: 10.1093/nar/4.11.3959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Groner Y., Gilboa E., Aviv H. Methylation and capping of RNA polymerase II primary transcripts by HeLa nuclear homogenates. Biochemistry. 1978 Mar 21;17(6):977–982. doi: 10.1021/bi00599a005. [DOI] [PubMed] [Google Scholar]
  19. Groner Y., Grosfeld H., Littauer U. Z. 5'-Capping structures of Artemia salina mRNA and the translational inhibition by cap analogs. Eur J Biochem. 1976 Dec;71(1):281–293. doi: 10.1111/j.1432-1033.1976.tb11114.x. [DOI] [PubMed] [Google Scholar]
  20. Groner Y., Hurwitz J. Synthesis of RNA containing a methylated blocked 5' terminus by HeLa nuclear homogenates. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2930–2934. doi: 10.1073/pnas.72.8.2930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Haegeman G., Fiers W. Evidence for 'splicing' of SV40 16S mRNA. Nature. 1978 May 4;273(5657):70–73. doi: 10.1038/273070a0. [DOI] [PubMed] [Google Scholar]
  22. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  23. Hsu M. T., Ford J. Sequence arrangement of the 5' ends of simian virus 40 16S and 19S mRNAs. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4982–4985. doi: 10.1073/pnas.74.11.4982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Khoury G., Carter B. J., Ferdinand F. J., Howley P. M., Brown M., Martin M. A. Genome localization of simian virus 40 RNA species. J Virol. 1976 Mar;17(3):832–840. doi: 10.1128/jvi.17.3.832-840.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lavi S., Groner Y. 5'-Terminal sequences and coding region of late simian virus 40 mRNAs are derived from noncontiguous segments of the viral genome. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5323–5327. doi: 10.1073/pnas.74.12.5323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Lavi U., Fernandez-Muñoz R., Darnell J. E., Jr Content of N-6 methyl adenylic acid in heterogeneous nuclear and messenger RNA of HeLa cells. Nucleic Acids Res. 1977 Jan;4(1):63–69. doi: 10.1093/nar/4.1.63. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. Reddy V. B., Thimmappaya B., Dhar R., Subramanian K. N., Zain B. S., Pan J., Ghosh P. K., Celma M. L., Weissman S. M. The genome of simian virus 40. Science. 1978 May 5;200(4341):494–502. doi: 10.1126/science.205947. [DOI] [PubMed] [Google Scholar]
  30. Revel M., Groner Y. Post-transcriptional and translational controls of gene expression in eukaryotes. Annu Rev Biochem. 1978;47:1079–1126. doi: 10.1146/annurev.bi.47.070178.005243. [DOI] [PubMed] [Google Scholar]
  31. Salditt-Georgieff M., Jelinek W., Darnell J. E., Furuichi Y., Morgan M., Shatkin A. Methyl labeling of HeLa cell hnRNA: a comparison with mRNA. Cell. 1976 Feb;7(2):227–237. doi: 10.1016/0092-8674(76)90022-2. [DOI] [PubMed] [Google Scholar]
  32. Schibler U., Kelley D. E., Perry R. P. Comparison of methylated sequences in messenger RNA and heterogeneous nuclear RNA from mouse L cells. J Mol Biol. 1977 Oct 5;115(4):695–714. doi: 10.1016/0022-2836(77)90110-3. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Sommer S., Salditt-Georgieff M., Bachenheimer S., Darnell J. E., Furuichi Y., Morgan M., Shatkin A. J. The methylation of adenovirus-specific nuclear and cytoplasmic RNA. Nucleic Acids Res. 1976 Mar;3(3):749–765. doi: 10.1093/nar/3.3.749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. 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]
  37. Wei C. M., Moss B. Nucleotide sequences at the N6-methyladenosine sites of HeLa cell messenger ribonucleic acid. Biochemistry. 1977 Apr 19;16(8):1672–1676. doi: 10.1021/bi00627a023. [DOI] [PubMed] [Google Scholar]
  38. Weinberg R. A., Warnaar S. O., Winocour E. Isolation and characterization of simian virus 40 ribonucleic acid. J Virol. 1972 Aug;10(2):193–201. doi: 10.1128/jvi.10.2.193-201.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES