Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1975 Apr;2(4):603–611. doi: 10.1093/nar/2.4.603

A rapid assay technique for RNA ribose methylases.

I Svensson, V J Patel
PMCID: PMC342867  PMID: 237257

Abstract

A rapid technique for quantitative separation of ribose-methylated nucleosides from base-methylated and non-methylated nucleosides by chromatography on DEAE-cellulose paper in the presence of borate is described. The method has been used as an assay for tRNA ribose methylases from yeast, using under methylated Escherichia coli tRNA as substrate. The main product formed with a partly purified yeast enzyme was characterized as 2'-O-methylcytidine.

Full text

PDF
603

Selected References

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

  1. Abbate J., Rottman F. Gas chromatographic method for determination of 2'-O-methylation in RNA. Anal Biochem. 1972 Jun;47(2):378–388. doi: 10.1016/0003-2697(72)90131-5. [DOI] [PubMed] [Google Scholar]
  2. Agris P. F., Spremulli L. L., Brown G. M. tRNA methylases from HeLa cells: purification and properties of an adenine-1-methylase and a guanine-N2-methylase. Arch Biochem Biophys. 1974 May;162(1):38–47. doi: 10.1016/0003-9861(74)90102-7. [DOI] [PubMed] [Google Scholar]
  3. Baskin F., Dekker C. A. A rapid and specific assay for sugar methylation in ribonucleic acid. J Biol Chem. 1967 Nov 25;242(22):5447–5449. [PubMed] [Google Scholar]
  4. Björk G. R., Isaksson L. A. Isolation of mutants of Escherichia coli lac king 5-methyluracil in transfer ribonucleic acid or 1-methylguanine in ribosomal RNA. J Mol Biol. 1970 Jul 14;51(1):83–100. doi: 10.1016/0022-2836(70)90272-x. [DOI] [PubMed] [Google Scholar]
  5. Dudock B. S., Katz G., Taylor E. K., Holley R. W. Primary structure of wheat germ phenylalanine transfer RNA. Proc Natl Acad Sci U S A. 1969 Mar;62(3):941–945. doi: 10.1073/pnas.62.3.941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gefter M. L. The in vitro synthesis of 2'-omethylguanosine and 2-methylthio 6N (gamma,gamma, dimethylallyl) adenosine in transfer RNA of Escherichia coli. Biochem Biophys Res Commun. 1969 Aug 7;36(3):435–441. doi: 10.1016/0006-291x(69)90583-x. [DOI] [PubMed] [Google Scholar]
  7. HALL R. H. ON THE 2'-O-METHYLRIBONUCLEOSIDE CONTENT OF RIBONUCLEIC ACIDS. Biochemistry. 1964 Jul;3:876–880. doi: 10.1021/bi00895a001. [DOI] [PubMed] [Google Scholar]
  8. Kerr S. J., Borek E. The tRNA methyltransferases. Adv Enzymol Relat Areas Mol Biol. 1972;36:1–27. doi: 10.1002/9780470122815.ch1. [DOI] [PubMed] [Google Scholar]
  9. Morisawa S., Chargaff E. On the bias of the distribution of the 2'-O-methylribonucleotide constituents of yeast transfer RNA. Biochim Biophys Acta. 1968 Dec 17;169(2):285–296. doi: 10.1016/0005-2787(68)90037-3. [DOI] [PubMed] [Google Scholar]
  10. Nichols J. L., Lane B. G. In vitro O2'-methylation of sugars in E. coli RNA. II. Methylation of ribosomal and transfer RNA by homologous methylases in crude cell-free extracts and particulate suspensions from a relaxed mutant of E. coli. Can J Biochem. 1968 Dec;46(12):1487–1495. doi: 10.1139/o68-222. [DOI] [PubMed] [Google Scholar]
  11. Nichols J. L., Lane B. G. N-4-methyl-2'-O-methyl cytidine and other methyl-substituted nucleoside constituents of Escherichia coli ribosomal and soluble RNA. Biochim Biophys Acta. 1966 Jun 22;119(3):649–651. doi: 10.1016/0005-2787(66)90147-x. [DOI] [PubMed] [Google Scholar]
  12. Phillips J. H., Kjellin-Stråby K. Studies on microbial ribonucleic acid. IV. Two mutants of Saccharomyces cerevisiae lacking N-2-dimethylguanine in soluble ribonucleic acid. J Mol Biol. 1967 Jun 28;26(3):509–518. doi: 10.1016/0022-2836(67)90318-x. [DOI] [PubMed] [Google Scholar]
  13. Pike L. M., Rottman F. The determination of 2'-O-methylnucleosides in RNA. Anal Biochem. 1974 Oct;61(2):367–378. doi: 10.1016/0003-2697(74)90404-7. [DOI] [PubMed] [Google Scholar]
  14. Rodeh R., Feldman M., Littauer U. Z. Properties of soluble ribonucleic acid methylases from rat liver. Biochemistry. 1967 Feb;6(2):451–460. doi: 10.1021/bi00854a013. [DOI] [PubMed] [Google Scholar]
  15. Starr J. L., Sells B. H. Methylated ribonucleic acids. Physiol Rev. 1969 Jul;49(3):623–669. doi: 10.1152/physrev.1969.49.3.623. [DOI] [PubMed] [Google Scholar]
  16. Svensson I., Björk G., Björk W., Johansson K. E., Johansson A. Evidence for enzymatic methylation in vitro of the ribose moiety of RNA. Biochem Biophys Res Commun. 1968 Apr 19;31(2):216–221. doi: 10.1016/0006-291x(68)90733-x. [DOI] [PubMed] [Google Scholar]
  17. Svensson I., Isaksson L., Henningsson A. Aminoacylation and polypeptide synthesis with tRNA lacking ribothymidine. Biochim Biophys Acta. 1971 May 13;238(2):331–337. doi: 10.1016/0005-2787(71)90100-6. [DOI] [PubMed] [Google Scholar]
  18. Svensson I. Studies on methionyl-tRNA synthetase. I. Effects of divalent and monovalent cations on methionyl-tRNA synthetase from Saccharomyces cerevisiae. Biochim Biophys Acta. 1967 Sep 12;146(1):239–252. doi: 10.1016/0005-2744(67)90090-3. [DOI] [PubMed] [Google Scholar]
  19. al-Arif A., Sporn M. B. An analytical method for the separation of sugar-methylated ribonucleosides from base-methylated and nonmethylated ribonucleosides. Anal Biochem. 1972 Aug;48(2):386–393. doi: 10.1016/0003-2697(72)90091-7. [DOI] [PubMed] [Google Scholar]

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

RESOURCES