Abstract
The effects of various polyanions including synthetic polynucleotides on DNApolymerases-alpha and -beta from blastulae of the sea urchin Hemicentrotus pulcherrimus and HeLa cells were studied. Only DNA polymerase-alpha was inhibited by polyanions, such as polyvinyl sufate, dextran sulfate, heparin, poly(G), poly(I), poly(U) and poly(ADP-Rib). Of the various polynucleotides tested, poly(G) and poly(I) were the strongest inhibitors. Kinetic studies showed that the Ki value for poly(G) was 0.3 microgram/ml and that poly(G) had 20-fold higher affinity than activated DNA for the template-primer site of DNA polymerase-alpha. Poly(U) and poly(ADP-Rib) were also inhibitory, but they were one hundredth as inhibitory as poly(G) or poly(I). Poly(A), poly(C), poly(A).poly(U) AND POLY(I).poly(C) were not inhibitory to DNA polymerase-alpha. In contrast, DNA olymerase-beta was not affected at all by these polyanions under the same conditions.
Full text
PDF











Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bertazzoni U., Stefanini M., Noy G. P., Giulotto E., Nuzzo F., Falaschi A., Spadari S. Variations of DNA polymerase-alpha and -beta during prolonged stimulation of human lymphocytes. Proc Natl Acad Sci U S A. 1976 Mar;73(3):785–789. doi: 10.1073/pnas.73.3.785. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bertazzoni U., Stefanini M., Pedrali-Noy G., Nuzzo F., Falaschi A. Levels of DNA polymerase-alpha and beta in normal and xeroderma pigmentosum fibroblasts. Nucleic Acids Res. 1977 Jan;4(1):141–148. doi: 10.1093/nar/4.1.141. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Brown D. G., Coffey D. S. Effects of polyinosinic acid and polycytidylic acid on the deoxyribonucleic acid template activity of isolated nuclei and soluble chromatin from rat liver. J Biol Chem. 1972 Dec 10;247(23):7674–7683. [PubMed] [Google Scholar]
- De Clercq E., Merigan T. C. Requirement of a stable secondary structure for the antiviral activity of polynucleotides. Nature. 1969 Jun 21;222(5199):1148–1152. doi: 10.1038/2221148a0. [DOI] [PubMed] [Google Scholar]
- Dicioccio R. A., Sahai Srivastava B. I. Structure-activity relationships and kinetic analyses of polyribonucleotide inhibition of human cellular deoxynucleotide-polymerizing enzymes. Biochim Biophys Acta. 1977 Oct 4;478(3):274–285. doi: 10.1016/0005-2787(77)90145-9. [DOI] [PubMed] [Google Scholar]
- Dube D. K., Seal G., Loeb L. A. Differential heat sensitivity of mammalian DNA polymerases. Biochem Biophys Res Commun. 1976 May 23;76(2):483–487. doi: 10.1016/0006-291x(77)90750-1. [DOI] [PubMed] [Google Scholar]
- Knopf K. W., Yamada M., Weissbach A. HeLa cell DNA polymerase gamma: further purification and properties of the enzyme. Biochemistry. 1976 Oct 5;15(20):4540–4548. doi: 10.1021/bi00665a032. [DOI] [PubMed] [Google Scholar]
- Loeb L. A. Purification and properties of deoxyribonucleic acid polymerase from nuclei of sea urchin embryos. J Biol Chem. 1969 Apr 10;244(7):1672–1681. [PubMed] [Google Scholar]
- Mosbaugh D. W., Stalker D. M., Probst G. S., Meyer R. R. Novikoff hepatoma deoxyribonucleic acid polymerase. Identification of a stimulatory protein bound to the beta-polymerase. Biochemistry. 1977 Apr 5;16(7):1512–1518. doi: 10.1021/bi00626a041. [DOI] [PubMed] [Google Scholar]
- Noy G. P., Weissbach A. HeLa cell DNA polymerases: the effect of cycloheximide in vivo and detection of a new form of DNA polymerase alpha. Biochim Biophys Acta. 1977 Jul 5;477(1):70–83. doi: 10.1016/0005-2787(77)90161-7. [DOI] [PubMed] [Google Scholar]
- Seki S., Oda T. Differential effects of heparin on replicative DNA synthesis and unscheduled DNA synthesis. Biochim Biophys Acta. 1977 Dec 14;479(4):391–399. doi: 10.1016/0005-2787(77)90032-6. [DOI] [PubMed] [Google Scholar]
- Sugimura T., Yoshimura N., Miwa M., Nagai H., Nagao M. Studies on poly(adenosine diphosphate-ribose). XI. Purification of poly(adenosine diphosphate-ribose) on a hydroxylapatite column. Arch Biochem Biophys. 1971 Dec;147(2):660–665. doi: 10.1016/0003-9861(71)90425-5. [DOI] [PubMed] [Google Scholar]
- Suzuki-Hori C., Nagano H., Mano Y. DNA polymerase-beta from the nuclear fraction of sea urchin embryos: characterization of the purified enzyme. J Biochem. 1977 Dec;82(6):1613–1621. doi: 10.1093/oxfordjournals.jbchem.a131857. [DOI] [PubMed] [Google Scholar]
- Wang TS-F, Fisher P. A., Sedwick W. D., Korn D. Identification of a new DNA polymerase activity in human KB cells. J Biol Chem. 1975 Jul 10;250(13):5270–5272. [PubMed] [Google Scholar]
- Weissbach A. Eukaryotic DNA polymerases. Annu Rev Biochem. 1977;46:25–47. doi: 10.1146/annurev.bi.46.070177.000325. [DOI] [PubMed] [Google Scholar]