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. 1984 Jun 25;12(12):5025–5036. doi: 10.1093/nar/12.12.5025

Chemical synthesis of 5-azacytidine nucleotides and preparation of tRNAs containing 5-azacytidine in its 3'-terminus.

W S Zielinski, M Sprinzl
PMCID: PMC318897  PMID: 6204276

Abstract

5-azacytidine-5'-triphosphate prepared from 5-azacytidine by chemical phosphorylation is a substrate for AMP (CMP) tRNA nucleotidyl transferase from yeast. tRNAsPhe from yeast containing 5-azacytidine in their 3'-termini were prepared enzymatically. tRNAPhe-Cpn5CpA and tRNAPhe-n5Cpn5CpA can be aminoacylated by phenylalanyl-tRNA synthetase from yeast and they are active in the poly(U)-dependent synthesis of poly(Phe) on E. coli ribosomes. The decomposition of 5-azacytidine via hydrolysis of the triazine ring is significantly accelerated by a phosphate group on the 5'-position of the nucleotide. After the incorporation of 5-azacytidine-5'-phosphate into a polynucleotide chain the rate of hydrolysis of the triazine ring decreases considerably.

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Selected References

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

  1. Bhanot O. S., Aoyagi S., Chambers R. W. Bisulfite-induced C changed to U transitions in yeast valine tRNA. J Biol Chem. 1977 Apr 25;252(8):2566–2574. [PubMed] [Google Scholar]
  2. Bhanot O. S., Chambers R. W. Bisulfite-induced C changed to U transitions in yeast alanine tRNA. J Biol Chem. 1977 Apr 25;252(8):2551–2559. [PubMed] [Google Scholar]
  3. HOARD D. E., OTT D. G. CONVERSION OF MONO- AND OLIGODEOXYRIBONUCLEOTIDES TO 5-TRIPHOSPHATES. J Am Chem Soc. 1965 Apr 20;87:1785–1788. doi: 10.1021/ja01086a031. [DOI] [PubMed] [Google Scholar]
  4. Jelenc P. C. Rapid purification of highly active ribosomes from Escherichia coli. Anal Biochem. 1980 Jul 1;105(2):369–374. doi: 10.1016/0003-2697(80)90472-8. [DOI] [PubMed] [Google Scholar]
  5. Jones P. A., Taylor S. M. Cellular differentiation, cytidine analogs and DNA methylation. Cell. 1980 May;20(1):85–93. doi: 10.1016/0092-8674(80)90237-8. [DOI] [PubMed] [Google Scholar]
  6. Lee T. T., Karon M. R. Inhibition of protein synthesis in 5-azacytidine-treated HeLa cells. Biochem Pharmacol. 1976 Aug 1;25(15):1737–1742. doi: 10.1016/0006-2952(76)90407-x. [DOI] [PubMed] [Google Scholar]
  7. Lee T. T., Momparler R. L. Enzymatic synthesis of 5-azacytidine 5'-triphosphate from 5-azacytidine. Anal Biochem. 1976 Mar;71(1):60–67. doi: 10.1016/0003-2697(76)90011-7. [DOI] [PubMed] [Google Scholar]
  8. Ley T. J., DeSimone J., Anagnou N. P., Keller G. H., Humphries R. K., Turner P. H., Young N. S., Keller P., Nienhuis A. W. 5-azacytidine selectively increases gamma-globin synthesis in a patient with beta+ thalassemia. N Engl J Med. 1982 Dec 9;307(24):1469–1475. doi: 10.1056/NEJM198212093072401. [DOI] [PubMed] [Google Scholar]
  9. McLaughlin L. W., Cramer F., Sprinzl M. Rapid analysis of modified tRNAphe from yeast by high-performance liquid chromatography: chromatography of oligonucleotides after RNase T1 digestion on aminopropylsilica and assignment of the fragments based on nucleoside analysis by chromatography on C18-silica. Anal Biochem. 1981 Mar 15;112(1):60–69. doi: 10.1016/0003-2697(81)90260-8. [DOI] [PubMed] [Google Scholar]
  10. Niwa O., Sugahara T. 5-Azacytidine induction of mouse endogenous type C virus and suppression of DNA methylation. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6290–6294. doi: 10.1073/pnas.78.10.6290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Notari R. E., DeYoung J. L. Kinetics and mechanisms of degradation of the antileukemic agent 5-azacytidine in aqueous solutions. J Pharm Sci. 1975 Jul;64(7):1148–1157. doi: 10.1002/jps.2600640704. [DOI] [PubMed] [Google Scholar]
  12. Randerath K., Tseng W. C., Harris J. S., Lu L. J. Specific effects of 5-fluoropyrimidines and 5-azapyrimidines on modification of the 5 position of pyrimidines, in particular the synthesis of 5-methyluracil and 5-methylcytosine in nucleic acids. Recent Results Cancer Res. 1983;84:283–297. doi: 10.1007/978-3-642-81947-6_22. [DOI] [PubMed] [Google Scholar]
  13. Reichman M., Penman S. The mechanism of inhibition of protein synthesis by 5-azacytidine in HeLa cells. Biochim Biophys Acta. 1973 Oct 12;324(2):282–289. doi: 10.1016/0005-2787(73)90145-7. [DOI] [PubMed] [Google Scholar]
  14. Riehl N., Remy P., Ebel J. P., Ehresmann B. Crosslinking of N-acetyl-phenylalanyl [s4U]tRNAPhe to protein S10 in the ribosomal P site. Eur J Biochem. 1982 Nov 15;128(2-3):427–433. doi: 10.1111/j.1432-1033.1982.tb06982.x. [DOI] [PubMed] [Google Scholar]
  15. Saiki J. H., McCredie K. B., Vietti T. J., Hewlett J. S., Morrison F. S., Costanzi J. J., Stuckey W. J., Whitecar J., Hoogstraten B. 5-azacytidine in acute leukemia. Cancer. 1978 Nov;42(5):2111–2114. doi: 10.1002/1097-0142(197811)42:5<2111::aid-cncr2820420505>3.0.co;2-i. [DOI] [PubMed] [Google Scholar]
  16. Schneider D., Solfert R., von der Haar F. Large scale purification of tRNA ser , tRNA tyr and tRNA phe from Baker's yeast. Hoppe Seylers Z Physiol Chem. 1972 Aug;353(8):1330–1336. doi: 10.1515/bchm2.1972.353.2.1330. [DOI] [PubMed] [Google Scholar]
  17. Schulman L. H., Goddard J. P. Loss of methionine acceptor activity resulting from a base change in the anticodon of Escherichia coli formylmethionine transfer ribonucleic acid. J Biol Chem. 1973 Feb 25;248(4):1341–1345. [PubMed] [Google Scholar]
  18. Sprinzl M., Cramer F. The -C-C-A end of tRNA and its role in protein biosynthesis. Prog Nucleic Acid Res Mol Biol. 1979;22:1–69. doi: 10.1016/s0079-6603(08)60798-9. [DOI] [PubMed] [Google Scholar]
  19. Sprinzl M., Sternbach H., von der Haar F., Cramer F. Enzymatic incorporation of ATP and CTP analogues into the 3' end of tRNA. Eur J Biochem. 1977 Dec;81(3):579–589. doi: 10.1111/j.1432-1033.1977.tb11985.x. [DOI] [PubMed] [Google Scholar]
  20. Sternbach H., von der Haar F., Schlimme E., Gaertner E., Cramer F. Isolation and properties of tRNA nucleotidyl transferase from yeast. Eur J Biochem. 1971 Sep 24;22(2):166–172. doi: 10.1111/j.1432-1033.1971.tb01528.x. [DOI] [PubMed] [Google Scholar]
  21. Thompson R. C., Karim A. M. The accuracy of protein biosynthesis is limited by its speed: high fidelity selection by ribosomes of aminoacyl-tRNA ternary complexes containing GTP[gamma S]. Proc Natl Acad Sci U S A. 1982 Aug;79(16):4922–4926. doi: 10.1073/pnas.79.16.4922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Venolia L., Gartler S. M., Wassman E. R., Yen P., Mohandas T., Shapiro L. J. Transformation with DNA from 5-azacytidine-reactivated X chromosomes. Proc Natl Acad Sci U S A. 1982 Apr;79(7):2352–2354. doi: 10.1073/pnas.79.7.2352. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wagner T., Sprinzl M. The complex formation between Escherichia coli aminoacyl-tRNA, elongation factor Tu and GTP. The effect of the side-chain of the amino acid linked to tRNA. Eur J Biochem. 1980;108(1):213–221. doi: 10.1111/j.1432-1033.1980.tb04714.x. [DOI] [PubMed] [Google Scholar]
  24. Yoshikawa M., Kato T., Takenishi T. A novel method for phosphorylation of nucleosides to 5'-nucleotides. Tetrahedron Lett. 1967 Dec;50:5065–5068. doi: 10.1016/s0040-4039(01)89915-9. [DOI] [PubMed] [Google Scholar]
  25. von der Haar F. Affinity elution as a purification method for aminoacyl-tRNA synthetases. Eur J Biochem. 1973 Apr 2;34(1):84–90. doi: 10.1111/j.1432-1033.1973.tb02731.x. [DOI] [PubMed] [Google Scholar]

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