Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1980 Jul 25;8(14):3175–3191. doi: 10.1093/nar/8.14.3175

Enzymatic synthesis of polyuridylic acid containing modified bases.

Y K Ho, J Aradi, T J Bardos
PMCID: PMC324363  PMID: 7443515

Abstract

5'Mercaptouridine-5'-diphosphate (hs5UDP) has been synthesized and investigated as a substrate of the polynucleotide phosphorylase of Micrococcus luteus. While hs5UDP is not utilized alone, it can be copolymerized with UDP; however, unusually for this enzyme, the ratio of 5'mercaptouridylate vs. uridylate residues in the polynucleotide product (MPU) is always lower than the ratio of hs5UDP v. UDP in the substrate mixture. Furthermore, hs5UDP decreases the rate of the enzymic polymerization reaction. The MPU product forms two-stranded and three-stranded complexes with poly(A). The circular dichroic spectra of these complexes are similar to those formed between poly(U) and poly(A), but their melting profiles indicate somewhat lower stability. The physicochemical and biochemical properties of the enzymic product are qualitatively similar to those of MPU prepared by chemical modification; both are potent inhibitors of a DNA-dependent RNA polymerase.

Full text

PDF
3175

Selected References

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

  1. Bardos T. J. Antimetabolites: molecular design and mode of action. Top Curr Chem. 1974;52:63–98. doi: 10.1007/3-540-06873-2_14. [DOI] [PubMed] [Google Scholar]
  2. Bardos T. J., Aradi J., Ho Y. K., Kalman T. I. Biochemical properties of 5-sulfur-substituted pyrimidine nucleosides and nucleotides. Ann N Y Acad Sci. 1975 Aug 8;255:522–531. doi: 10.1111/j.1749-6632.1975.tb29255.x. [DOI] [PubMed] [Google Scholar]
  3. Bardos T. J., Kalman T. I. Spectrophotometric and chemical studies of 5-mercaptouracil, 5-mercaptodeoxyuridine, and their S-substituted derivatives. J Pharm Sci. 1966 Jun;55(6):606–610. doi: 10.1002/jps.2600550615. [DOI] [PubMed] [Google Scholar]
  4. Bähr W., Faerber P., Scheit K. H. The effects of thioketo substitution upon uracil-adenine interactions in polyribonucleotides. Synthesis and properties of poly (2-thiouridylic acid) and poly(2,4-dithiouridylic acid). Eur J Biochem. 1973 Mar 15;33(3):535–544. doi: 10.1111/j.1432-1033.1973.tb02713.x. [DOI] [PubMed] [Google Scholar]
  5. Chandra P., Bardos T. J. Inhibition of DNA polymerases from RNA tumor viruses by novel template analogues: partially thiolated polycytidylic acid. Res Commun Chem Pathol Pharmacol. 1972 Nov;4(3):615–622. [PubMed] [Google Scholar]
  6. Chandra P., Ebener U., Bardos T. J., Chakrabarti P., Ho Y. K., Mikulski A. J., Zsindely A. Polynucleotides containing 5-mercapto-substituted pyrimidines: inhibition of viral DNA polymerases and the biological implication. Ann N Y Acad Sci. 1975 Aug 8;255:532–543. doi: 10.1111/j.1749-6632.1975.tb29256.x. [DOI] [PubMed] [Google Scholar]
  7. Chandra P., Ebener U., Steel L. K., Laube H., Gericke D., Mildner B., Bardos T. J., Ho Y. K., Götz A. Molecular approaches to inhibit oncogenesis by RNA tumor viruses. Ann N Y Acad Sci. 1977 Mar 4;284:444–462. doi: 10.1111/j.1749-6632.1977.tb21981.x. [DOI] [PubMed] [Google Scholar]
  8. Eaton M. A., Hutchinson D. W. Poly(5-chlorocytidylic acid). Biochemistry. 1972 Aug 15;11(17):3162–3167. doi: 10.1021/bi00767a004. [DOI] [PubMed] [Google Scholar]
  9. Eaton M. A., Hutchinson D. W. Poly(5-hydroxycytidylic acid) . Biochim Biophys Acta. 1973 Sep 7;319(3):281–287. doi: 10.1016/0005-2787(73)90167-6. [DOI] [PubMed] [Google Scholar]
  10. FELSENFELD G., RICH A. Studies on the formation of two- and three-stranded polyribonucleotides. Biochim Biophys Acta. 1957 Dec;26(3):457–468. doi: 10.1016/0006-3002(57)90091-4. [DOI] [PubMed] [Google Scholar]
  11. Ho Y. K., Fiel R. J., Aradi J., Bardos T. J. Structural characterization of partially thiolated poly(cytidylic acid). Biochemistry. 1979 Dec 11;18(25):5630–5635. doi: 10.1021/bi00592a017. [DOI] [PubMed] [Google Scholar]
  12. Ho Y. K., Kung M. P., Bardos T. J. Oligonucleotides containing modified bases. I. Inhibition of DNA and RNA polymerases by partially thiolated oligocytidylic acids. Biochem Biophys Res Commun. 1976 Dec 20;73(4):903–910. doi: 10.1016/0006-291x(76)90207-2. [DOI] [PubMed] [Google Scholar]
  13. Ho Y. K., Preisler H. D., Bardos T. J. Effects of partially thiolated polycytidylic acid on the clonogenicity of murine leukemic stem cells. Cancer Res. 1979 Aug;39(8):3163–3166. [PubMed] [Google Scholar]
  14. Ho Y. K. Studies on the mode of action of partially thiolated polycytidylic acid (MPC), a novel type of antineoplastic agent. Res Commun Chem Pathol Pharmacol. 1979 Aug;25(2):293–306. [PubMed] [Google Scholar]
  15. Howard F. B., Frazier J., Miles H. T. Interaction of poly-5-bromocytidylic acid with polyinosinic acid. A study of helix stability and spectroscopic properties. J Biol Chem. 1969 Mar 10;244(5):1291–1302. [PubMed] [Google Scholar]
  16. Kröger M., Singer B. Influence of different levels of 2-thiocytidine on physical and template properties of cytidine--2-thiocytidine copolymers. Biochemistry. 1979 Jan 9;18(1):91–95. doi: 10.1021/bi00568a014. [DOI] [PubMed] [Google Scholar]
  17. Kung M. P., Ho Y. K., Bardos T. J. Action of partially thiolated polynucleotides on the DNA polymerase alpha from regenerating rat liver. Cancer Res. 1976 Dec;36(12):4537–4542. [PubMed] [Google Scholar]
  18. Michelson A. M., Monny C. Polynucleotide analogues. XII. Poly 5-bromocytidylic acid and poly 5-iodocytidylic acid. Biochim Biophys Acta. 1967 Nov 21;149(1):88–98. doi: 10.1016/0005-2787(67)90693-4. [DOI] [PubMed] [Google Scholar]
  19. Mikulski A. J., Bardos T. J., Chakrabarti P., Kalman T. I., Zsindely A. Inhibition of DNA-dependent RNA polymerase with partially thiolated polynucleotides. Biochim Biophys Acta. 1973 Sep 7;319(3):294–303. doi: 10.1016/0005-2787(73)90169-x. [DOI] [PubMed] [Google Scholar]
  20. Miller W. L., Brenner D. J., Doctor B. P. Fractionation and molecular weight determination of deoxyribonucleic acid fragments using agarose column chromatography. Anal Biochem. 1974 Oct;61(2):454–463. doi: 10.1016/0003-2697(74)90412-6. [DOI] [PubMed] [Google Scholar]
  21. O'Malley J. A., Ho Y. K., Chakrabarti P., DiBerardino L., Chandra P., Orinda D. A., Byrd D. M., Bardos T. J., Carter W. A. Antiviral activity of partially thiolated polynucleotides. Mol Pharmacol. 1975 Jan;11(1):61–69. [PubMed] [Google Scholar]
  22. OCHOA S., MII S. Enzymatic synthesis of polynucleotides. IV. Purification and properties of polynucleotide phosphorylase of Azotobacter vinelandii. J Biol Chem. 1961 Dec;236:3303–3311. [PubMed] [Google Scholar]
  23. SINGER M. F., GUSS J. K. The dependence of reactions catalyzed by polynucleotide phosphorylase on oligonucleotides. J Biol Chem. 1962 Jan;237:182–189. [PubMed] [Google Scholar]
  24. Scheit K. H., Gaertner E. Die Polymerisation von 4-Thiouridin-5'-diphosphat und 4-thiothymidin-5'-diphosphat durch Polynucleotidphosphorylase aus Micrococcus lysodeikticus. Biochim Biophys Acta. 1969 May 20;182(1):1–9. [PubMed] [Google Scholar]
  25. Simuth J., Scheit K. H., Gottschalk E. M. The enzymatic synthesis of poly 4-thiouridylic acid by polynucleotide phosphorylase from Escherichia coli. Biochim Biophys Acta. 1970 Apr 15;204(2):371–380. doi: 10.1016/0005-2787(70)90156-5. [DOI] [PubMed] [Google Scholar]
  26. Srivastava B. I., Bardos T. J. Inhibition of some DNA polymerase activities from cultured Burkitt cells by thiolated ribonucleic acids. Life Sci. 1973 Jul 1;13(1):47–53. doi: 10.1016/0024-3205(73)90276-2. [DOI] [PubMed] [Google Scholar]
  27. Swierkowski M., Shugar D. Poly 5-ethyluridylic acid, a polyuridylic acid analogue. J Mol Biol. 1970 Jan 14;47(1):57–67. doi: 10.1016/0022-2836(70)90401-8. [DOI] [PubMed] [Google Scholar]
  28. Szabo L., Kalman T. I., Bardos T. J. The reaction of 5-bromouracil derivatives with sulfur nucleophiles, and a novel synthetic route to 5-sulfur-substituted uracils and nucleotides. J Org Chem. 1970 May;35(5):1434–1437. doi: 10.1021/jo00830a038. [DOI] [PubMed] [Google Scholar]

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

RESOURCES