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. 1975 Oct;2(10):1653–1668. doi: 10.1093/nar/2.10.1653

The methylation state of poly A-containing messenger RNA from cultured hamster cells.

D T Dubin, R H Taylor
PMCID: PMC343535  PMID: 1187339

Abstract

The poly A-containing mRNA of cultured hamster (BHK-21) cells has been examined with regard to methylation status. Steady state-labeled mRNA was obtained by incubating cells for 20-22h in the presence of [methyl-3H]-methionine and 32Pi. The degree of methylation of this RNA was 1.8 methyl groups per 1000 nucleotides, or 4-5 methyl groups on the average per molecule. The nature of the methylated residues was determined by paper chromatography and electrophoresis of acid and alkaline hydrolysates, by DEAE cellulose chromatography of alkaline hydrolysates and of T2 RNase digests, and by examining the effect of subjecting samples to "beta-elimination." Approx. half of the methyl groups occurred in standard ("internal") linkage, 10% as m5Cp and 40% as m6Ap residues. The remainder occurred at least for the most part in "blocked" 5'-termini with the presumptive structure m7G(5')ppp(Nm)p.., where Nm was Gm, m6Am, Um, or Cm.

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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. Desrosiers R., Friderici K., Rottman F. Identification of methylated nucleosides in messenger RNA from Novikoff hepatoma cells. Proc Natl Acad Sci U S A. 1974 Oct;71(10):3971–3975. doi: 10.1073/pnas.71.10.3971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dubin D. T. Methylated nucleotide content of mitochondrial ribosomal RNA from hamster cells. J Mol Biol. 1974 Apr 5;84(2):257–273. doi: 10.1016/0022-2836(74)90584-1. [DOI] [PubMed] [Google Scholar]
  4. Dubin D. T., Montenecourt B. S. Mitochondrial RNA from cultured animal cells. Distinctive high-molecular-weight and 4 s species. J Mol Biol. 1970 Mar 14;48(2):279–295. doi: 10.1016/0022-2836(70)90161-0. [DOI] [PubMed] [Google Scholar]
  5. Faust M., Millward S. In vitro methylation of nascent reovirus mRNA by a virion-associated methyl transferase. Nucleic Acids Res. 1974 Dec;1(12):1739–1752. doi: 10.1093/nar/1.12.1739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. Furuichi Y., Morgan M., Shatkin A. J., Jelinek W., Salditt-Georgieff M., Darnell J. E. Methylated, blocked 5 termini in HeLa cell mRNA. Proc Natl Acad Sci U S A. 1975 May;72(5):1904–1908. doi: 10.1073/pnas.72.5.1904. [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. Hirsch M., Penman S. The messenger-like properties of the poly(A)plus RNA in mammalian mitochondria. Cell. 1974 Dec;3(4):335–339. doi: 10.1016/0092-8674(74)90047-6. [DOI] [PubMed] [Google Scholar]
  11. LITTLEFIELD J. W., DUNN D. B. The occurrence and distribution of thymine and three methylated-adenine bases in ribonucleic acids from several sources. Biochem J. 1958 Dec;70(4):642–651. doi: 10.1042/bj0700642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Maden B. E., Salim M. The methylated nucleotide sequences in HELA cell ribosomal RNA and its precursors. J Mol Biol. 1974 Sep 5;88(1):133–152. doi: 10.1016/0022-2836(74)90299-x. [DOI] [PubMed] [Google Scholar]
  13. Muthukrishnan S., Both G. W., Furuichi Y., Shatkin A. J. 5'-Terminal 7-methylguanosine in eukaryotic mRNA is required for translation. Nature. 1975 May 1;255(5503):33–37. doi: 10.1038/255033a0. [DOI] [PubMed] [Google Scholar]
  14. Perry R. P., Kelley D. E. Messenger RNA turnover in mouse L cells. J Mol Biol. 1973 Oct 5;79(4):681–696. doi: 10.1016/0022-2836(73)90071-5. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Roy P., Bishop D. H. Initiation and direction of RNA transcription by vesicular stomatitis virus virion transcriptase. J Virol. 1973 Apr;11(4):487–501. doi: 10.1128/jvi.11.4.487-501.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Singer R. H., Penman S. Messenger RNA in HeLa cells: kinetics of formation and decay. J Mol Biol. 1973 Aug 5;78(2):321–334. doi: 10.1016/0022-2836(73)90119-8. [DOI] [PubMed] [Google Scholar]
  18. 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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