Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1986 Feb 11;14(3):1131–1148. doi: 10.1093/nar/14.3.1131

A single tRNA (guanine)-methyltransferase from Tetrahymena with both mono- and di-methylating activity.

M P Reinhart, J M Lewis, P S Leboy
PMCID: PMC339493  PMID: 3081876

Abstract

A tRNA (guanine-2) methyltransferase has been purified to homogeneity from the protozoan Tetrahymena pyriformis. The enzyme methylates purified E. coli tRNAs which have a guanine residue at position 26 from the 5' end; it also methylates tRNA prepared from the m22G- yeast mutant trm 1. This methyltransferase is therefore equivalent to the guanine methyltransferase 2mGII found in mammalian extracts. The purified 2mGII from Tetrahymena is capable of forming both N2-methylguanine and N22-dimethylguanine on a single tRNA isoaccepting species; under conditions of limiting tRNA or long reaction times the predominant product is dimethylguanine. Analysis of the products formed under varying reaction conditions suggests that dimethylguanine formation is a two step process requiring dissociation of the enzyme-monomethylated tRNA intermediate.

Full text

PDF
1135

Images in this article

Selected References

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

  1. 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]
  2. Andrews P. Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochem J. 1964 May;91(2):222–233. doi: 10.1042/bj0910222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brunke K. J., Leboy P. S. An unusual transfer RNA (guanine-2-)-methyltransferase from transplantable rat mammary tumors. Cancer Res. 1982 Dec;42(12):4979–4984. [PubMed] [Google Scholar]
  4. Bryan J. K. Molecular weights of protein multimers from polyacrylamide gel electrophoresis. Anal Biochem. 1977 Apr;78(2):513–519. doi: 10.1016/0003-2697(77)90111-7. [DOI] [PubMed] [Google Scholar]
  5. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  6. FERGUSON K. A. STARCH-GEL ELECTROPHORESIS--APPLICATION TO THE CLASSIFICATION OF PITUITARY PROTEINS AND POLYPEPTIDES. Metabolism. 1964 Oct;13:SUPPL–SUPPL1002. doi: 10.1016/s0026-0495(64)80018-4. [DOI] [PubMed] [Google Scholar]
  7. Ginell S. L., Parthasarathy R. Conformation of N2-methylguanosine, a modified nucleoside of tRNA. Biochem Biophys Res Commun. 1978 Oct 30;84(4):886–894. doi: 10.1016/0006-291x(78)91666-2. [DOI] [PubMed] [Google Scholar]
  8. Glick J. M., Averyhart V. M., Leboy P. S. Purification and characterization of two tRNA-(guanine)-methyltransferases from rat liver. Biochim Biophys Acta. 1978 Mar 29;518(1):158–171. doi: 10.1016/0005-2787(78)90125-9. [DOI] [PubMed] [Google Scholar]
  9. Glick J. M., Leboy P. S. Purification and properties of tRNA(adenine-1)-methyltransferase from rat liver. J Biol Chem. 1977 Jul 25;252(14):4790–4795. [PubMed] [Google Scholar]
  10. Greenberg R., Dudock B. Isolation and chracterization of m5U-methyltransferase from Escherichia coli. J Biol Chem. 1980 Sep 10;255(17):8296–8302. [PubMed] [Google Scholar]
  11. Keith J. M., Winters E. M., Moss B. Purification and characterization of a HeLa cell transfer RNA(cytosine-5-)-methyltransferase. J Biol Chem. 1980 May 25;255(10):4636–4644. [PubMed] [Google Scholar]
  12. Kersten H. Alteration of tRNA modification in eukaryotes: causes and consequences. Recent Results Cancer Res. 1983;84:255–263. doi: 10.1007/978-3-642-81947-6_19. [DOI] [PubMed] [Google Scholar]
  13. Kraus J., Staehelin M. N2-guanine specific transfer RNA methyltransferase II from rat liver. Nucleic Acids Res. 1974 Nov;1(11):1479–1496. doi: 10.1093/nar/1.11.1479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kuchino Y., Nishimura S. Nucleotide sequence specificities of guanylate residue-specific tRNA methylases from rat liver. Biochem Biophys Res Commun. 1970 Jul 27;40(2):306–313. doi: 10.1016/0006-291x(70)91010-7. [DOI] [PubMed] [Google Scholar]
  15. Marcu K. B., Dudock B. S. Effect of ribothymidine in specific eukaryotic tRNAs on their efficiency in in vitro protein synthesis. Nature. 1976 May 13;261(5556):159–162. doi: 10.1038/261159a0. [DOI] [PubMed] [Google Scholar]
  16. Pegg A. E. Sites of methylation of purified transfer ribonucleic acid preparations by enzymes from normal tissues and from tumours induced by dimethylnitrosamine and 1,2-dimethylhydrazine. Biochem J. 1974 Feb;137(2):239–248. doi: 10.1042/bj1370239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Roe B. A., Tsen H. Y. Role of ribothymidine in mammalian tRNAPhe. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3696–3700. doi: 10.1073/pnas.74.9.3696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Roe B., Michael M., Dudock B. Function of N2 methylguanine in phenylalanine transfer RNA. Nat New Biol. 1973 Dec 5;246(153):135–138. doi: 10.1038/newbio246135a0. [DOI] [PubMed] [Google Scholar]
  20. Salas C. E., Dirheimer G. In vitro methylation of yeast tRNAAsp by rat brain cortical tRNA-(adenine-1) methyltransferase. Nucleic Acids Res. 1979 Mar;6(3):1123–1133. doi: 10.1093/nar/6.3.1123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schaffner W., Weissmann C. A rapid, sensitive, and specific method for the determination of protein in dilute solution. Anal Biochem. 1973 Dec;56(2):502–514. doi: 10.1016/0003-2697(73)90217-0. [DOI] [PubMed] [Google Scholar]
  22. Spremulli L. L., Agris P. F., Brown G. M., Rajbhandary U. L. Escherichia coli formylmethionine tRNA: methylation of specific guanine and adenine residues catalyzed by HeLa cells tRNA methylases and the effect of these methylations on its biological properties. Arch Biochem Biophys. 1974 May;162(1):22–37. doi: 10.1016/0003-9861(74)90101-5. [DOI] [PubMed] [Google Scholar]

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

RESOURCES