Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1987 May 26;15(10):3997–4006. doi: 10.1093/nar/15.10.3997

Template-primer analogs as substrates for DNA polymerase.

T R Webb, P Jhurani, P G Ng
PMCID: PMC340827  PMID: 2438649

Abstract

In order to gain more understanding about the mode of action of DNA polymerase, eight related partially self-complementary "hairpin" shaped oligodeoxynucleotides were prepared. Four of the oligomers contained either 1-beta-D-arabinofuranosyluracil (ara-U) or 1-beta-D-2'deoxyxylofuranosylthymine (dxT) nucleoside analogs at their 3' termini. We investigated the ability of the oligomers to prime DNA synthesis in relationship to the stability of the hybridized region, the nature of the sugar terminus and the DNA polymerase used (reverse transcriptase, or polymerase alpha). The results are discussed in relation to the mode of action of some nucleoside analog inhibitors of DNA polymerase. An understanding of the mechanism of DNA polymerase action was used to design template-primer analogs as polymerase inhibitors. Two of the oligodeoxynucleotide analogs prepared were found to be potent inhibitors of polymerase alpha.

Full text

PDF
3997

Images in this article

3999Image
on p.3999
4001Image
on p.4001

Selected References

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

  1. Atkinson M. R., Deutscher M. P., Kornberg A., Russell A. F., Moffatt J. G. Enzymatic synthesis of deoxyribonucleic acid. XXXIV. Termination of chain growth by a 2',3'-dideoxyribonucleotide. Biochemistry. 1969 Dec;8(12):4897–4904. doi: 10.1021/bi00840a037. [DOI] [PubMed] [Google Scholar]
  2. Caruthers M. H. Gene synthesis machines: DNA chemistry and its uses. Science. 1985 Oct 18;230(4723):281–285. doi: 10.1126/science.3863253. [DOI] [PubMed] [Google Scholar]
  3. Chow F., Kempe T., Palm G. Synthesis of oligodeoxyribonucleotides on silica gel support. Nucleic Acids Res. 1981 Jun 25;9(12):2807–2817. doi: 10.1093/nar/9.12.2807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Derse D., Cheng Y. C., Furman P. A., St Clair M. H., Elion G. B. Inhibition of purified human and herpes simplex virus-induced DNA polymerases by 9-(2-hydroxyethoxymethyl)guanine triphosphate. Effects on primer-template function. J Biol Chem. 1981 Nov 25;256(22):11447–11451. [PubMed] [Google Scholar]
  5. Fisher P. A., Korn D. Enzymological characterization of KB cell DNA polymerase-alpha. II. Specificity of the protein-nucleic acid interaction. J Biol Chem. 1979 Nov 10;254(21):11033–11039. [PubMed] [Google Scholar]
  6. Froehler B. C., Ng P. G., Matteucci M. D. Synthesis of DNA via deoxynucleoside H-phosphonate intermediates. Nucleic Acids Res. 1986 Jul 11;14(13):5399–5407. doi: 10.1093/nar/14.13.5399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Haasnoot C. A., den Hartog J. H., de Rooij J. F., van Boom J. H., Altona C. Loopstructures in synthetic oligodeoxynucleotides. Nucleic Acids Res. 1980 Jan 11;8(1):169–181. doi: 10.1093/nar/8.1.169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ikegami S., Taguchi T., Ohashi M., Oguro M., Nagano H., Mano Y. Aphidicolin prevents mitotic cell division by interfering with the activity of DNA polymerase-alpha. Nature. 1978 Oct 5;275(5679):458–460. doi: 10.1038/275458a0. [DOI] [PubMed] [Google Scholar]
  9. Ikuta S., Chattopadhyaya R., Ito H., Dickerson R. E., Kearns D. R. NMR study of a synthetic DNA hairpin. Biochemistry. 1986 Aug 26;25(17):4840–4849. doi: 10.1021/bi00365a018. [DOI] [PubMed] [Google Scholar]
  10. Kufe D. W., Major P. P. Studies on the mechanism of action of cytosine arabinoside. Med Pediatr Oncol. 1982;10 (Suppl 1):49–67. doi: 10.1002/mpo.2950100708. [DOI] [PubMed] [Google Scholar]
  11. McMurry J. E., Webb T. R. Synthesis of a tricyclic aphidicolin analogue that inhibits DNA synthesis in vitro. J Med Chem. 1984 Oct;27(10):1367–1369. doi: 10.1021/jm00376a028. [DOI] [PubMed] [Google Scholar]
  12. Momparler R. L. Kinetic and template studies with 1- -D-arabinofuranosylcytosine 5'-triphosphate and mammalian deoxyribonucleic acid polymerase. Mol Pharmacol. 1972 May;8(3):362–370. [PubMed] [Google Scholar]
  13. Verma I. M. The reverse transcriptase. Biochim Biophys Acta. 1977 Mar 21;473(1):1–38. doi: 10.1016/0304-419x(77)90005-1. [DOI] [PubMed] [Google Scholar]
  14. Wilson S. H., Matsukage A., Bohn E. W., Chen Y. C., Sivarajan M. Polynucleotide recognition by DNA alpha-polymerase. Nucleic Acids Res. 1977 Nov;4(11):3981–3996. doi: 10.1093/nar/4.11.3981. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES