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. 1976 Jun;3(6):1603–1611. doi: 10.1093/nar/3.6.1603

Poly 2'-O-ethylcytidylate, an inhibitor and poor template for AMV reverse transcriptase.

R Mikke, M Kielanowska, D Shugar, B Zmudzka
PMCID: PMC343017  PMID: 60742

Abstract

Poly(2'-O-ethylcytidylate) is a poor template-primer for purified avian myeloblastosis virus reverse transcriptase; the relative activities of the template-primers poly(C)-oligo(dG), poly(Cm)-oligo(dG) and poly(Ce)-oligo(dG) are 23:16:1. A mixture of poly(Ce) and poly(dI) is inactive as template-primer, in agreement with the observed inability of these to form a helical complex. By contrast the inactivity of poly(Ce)-poly(I) is shown to be due to the influence of the 2'-O-ethyl residue. Poly(Ce) inhibits poly(A)-oligo(dT)-directed polymerase activity, with Ki = 3 muM, but marked inhibition with poly(A)-poly(dT) occurs only at low concentrations of the latter. Poly(Ce) did not inhibit template-primer activity of poly(C)-poly(dI) and poly(dC)-poly(dI). Qualitative physico-chemical studies show only partial complex formation between oligo(dG) and poly(C) and its 2'-O-alkyl analogues. This is discussed in relation to the widespread use of poly(C)-oligo(dG) as the template-primer for reverse transcriptase.

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Selected References

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

  1. Abrell J. W., Gallo R. C. Purification, characterization, and comparison of the DNA polymerases from two primate RNA tumor viruses. J Virol. 1973 Sep;12(3):431–439. doi: 10.1128/jvi.12.3.431-439.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arya S. K., Carter W. A., Alderfer J. L., Ts'o P. O. Inhibition of RNA directed DNA polymerase of murine leukemia virus by 2'-O-alkylated polyadenylic acids. Biochem Biophys Res Commun. 1974 Jul 24;59(2):608–615. doi: 10.1016/s0006-291x(74)80023-9. [DOI] [PubMed] [Google Scholar]
  3. Bollum F. J. Mammalian DNA polymerases. Prog Nucleic Acid Res Mol Biol. 1975;15(0):109–144. doi: 10.1016/s0079-6603(08)60118-x. [DOI] [PubMed] [Google Scholar]
  4. CHAMBERLIN M. J., PATTERSON D. L. PHYSICAL AND CHEMICAL CHARACTERIZATION OF THE ORDERED COMPLEXES FORMED BETWEEN POLYINOSINIC ACID, POLYCYTIDYLIC ACID AND THEIR DEOXYRIBO-ANALOGUES. J Mol Biol. 1965 Jun;12:410–428. doi: 10.1016/s0022-2836(65)80264-9. [DOI] [PubMed] [Google Scholar]
  5. De Clercq E., Billiau A., Hattori M., Ikehara M. Inhibition of oncornavirus functions by poly (2-methylthioinosinic acid). Nucleic Acids Res. 1975 Dec;2(12):2305–2313. doi: 10.1093/nar/2.12.2305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. De Clercq E., Billiau A., Hobbs J., Torrence P. F., Witkop B. Inhibition of oncornavirus functions by 2'-azido polynucleotides. Proc Natl Acad Sci U S A. 1975 Jan;72(1):284–288. doi: 10.1073/pnas.72.1.284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. De Clercq E., Zmudzka B., Shugar D. Antiviral activity of polynucleotides: role of the 2'-hydroxyl and a pyrimidine 5-methyl. FEBS Lett. 1972 Jul 15;24(1):137–140. doi: 10.1016/0014-5793(72)80845-7. [DOI] [PubMed] [Google Scholar]
  8. Erickson R. J., Grosch J. C. The inhibition of avian myeloblastosis virus deoxyribonucleic acid polymerase by synthetic polynucleotides. Biochemistry. 1974 Apr 23;13(9):1987–1993. doi: 10.1021/bi00706a032. [DOI] [PubMed] [Google Scholar]
  9. Erickson R. J., Janik B., Sommer R. G. The inhibition of the avian myeloblastosis virus DNA polymerase by poly(U) fractions of varying chain length. Biochem Biophys Res Commun. 1973 Jun 19;52(4):1475–1482. doi: 10.1016/0006-291x(73)90667-0. [DOI] [PubMed] [Google Scholar]
  10. Gerard G. F., Loewenstein P. M., Green M., Rottman F. Detection of reverse transcriptase in human breast tumours with poly(Cm)-oligo(dG). Nature. 1975 Jul 10;256(5513):140–143. doi: 10.1038/256140a0. [DOI] [PubMed] [Google Scholar]
  11. Gerard G. F. Poly (2'O-methylcytidylate).oligodeoxyguanylate, a template-primer specific for reverse transcriptase, is not utilized by HeLa cell gamma DNA polymerases. Biochem Biophys Res Commun. 1975 Apr 7;63(3):706–711. doi: 10.1016/s0006-291x(75)80441-4. [DOI] [PubMed] [Google Scholar]
  12. Gerard G. F., Rottman F., Green M. Poly(2'-O-methylcytidylate)-oligodeoxyguanylate as a template for the ribonucleic acid directed deoxyribonucleic acid polymerase in ribonucleic acid tumor virus particles and a specific probe for the ribonucleic acid directed enzyme in transformed murine cells. Biochemistry. 1974 Apr 9;13(8):1632–1641. doi: 10.1021/bi00705a012. [DOI] [PubMed] [Google Scholar]
  13. Kacian D. L., Spiegelman S. Purification and detection of reverse transcriptase in viruses and cells. Methods Enzymol. 1974;29:150–173. doi: 10.1016/0076-6879(74)29018-9. [DOI] [PubMed] [Google Scholar]
  14. Lefler C. F., Bollum F. J. Deoxynucleotide-polymerizing enzymes of calf thymus gland. 3. Preparation of poly N-acetyldeoxyguanylate and polydeoxyguanylate. J Biol Chem. 1969 Feb 25;244(4):594–601. [PubMed] [Google Scholar]
  15. Leis J. P., Berkower I., Hurwitz J. Mechanism of action of ribonuclease H isolated from avian myeloblastosis virus and Escherichia coli. Proc Natl Acad Sci U S A. 1973 Feb;70(2):466–470. doi: 10.1073/pnas.70.2.466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Siedlecki J. A., Zmudzka B. Role of Mn2+ in the reaction of polynucleotide phosphorylase with 2'-O-methylated substrates. Acta Biochim Pol. 1975;22(2):163–168. [PubMed] [Google Scholar]
  17. Spiegelman S., Burny A., Das M. R., Keydar J., Schlom J., Trávnícek M., Watson K. Synthetic DNA-RNA hybrids and RNA-RNA duplexes as templates for the polymerases of the oncogenic RNA viruses. Nature. 1970 Oct 31;228(5270):430–432. doi: 10.1038/228430a0. [DOI] [PubMed] [Google Scholar]
  18. Srivastava B. I. Modified nucleotide polymers as inhibitors of DNA polymerases. Biochim Biophys Acta. 1975 Dec 4;414(2):126–132. doi: 10.1016/0005-2787(75)90215-4. [DOI] [PubMed] [Google Scholar]
  19. Tuominen F. W., Kenney F. T. Inhibition of the DNA polymerase of Rauscher leukemia virus by single-stranded polyribonucleotides. Proc Natl Acad Sci U S A. 1971 Sep;68(9):2198–2202. doi: 10.1073/pnas.68.9.2198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Zmudzka B., Janion C., Shugar D. Poly 2'-O-methylcytidylic acid and the role of the 2'-hydroxyl in polynucleotide structure. Biochem Biophys Res Commun. 1969 Dec 4;37(6):895–901. doi: 10.1016/0006-291x(69)90215-0. [DOI] [PubMed] [Google Scholar]
  21. Zmudzka B., Tichy M., Shugar D. The structure of poly 2'-O-methylcytidylic acid and its complexes with polyinosinic acid. Acta Biochim Pol. 1972;19(2):149–160. [PubMed] [Google Scholar]

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