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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1980 Jun;77(6):3317–3321. doi: 10.1073/pnas.77.6.3317

A beta-like DNA polymerase activity in the slime mold Dictyostelium discoideum.

E F Baril, C Scheiner, T Pederson
PMCID: PMC349606  PMID: 6774334

Abstract

Two distinct DNA polymerases (DNA nucleotidyltransferases) have been partially purified from exponentially growing cells of the slime mold Dictyostelium discoideum. The major DNA polymerase activity, DNA polymerase A, has physical and enzymatic properties similar to DNA polymerase alpha from vertebrate cells. DNA polymerase B, representing less than 10% of the polymerase activity in homogenates, is similar to DNA polymerase beta from vertebrate cells in that its activity is not inhibited by N-ethylmaleimide, aphidicolin, or KCl at a concentration below 200 mM. Like DNA polymerase beta, the activity of Dictyostelium DNA polymerase B is sensitive to 2', 3'-dideoxythymidine 5'-triphosphate. However, it differs from vertebrate DNA polymerase beta in that it chromatographs as an acidic protein and has an apparent molecular weight of 70,000. Previous reports of the absence of a beta-like DNA polymerase in D. discoideum apparently resulted from inadequate extraction and assay conditions for the enzyme. It seems quite probable from the results of this study and those of other studies (e.g., yeast and Tetrahymena pyriformis) that lower eukaryotic organisms, as well as vertebrates and prokaryotes, contain more than one form of DNA polymerase.

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Selected References

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

  1. BOLLUM F. J. Calf thymus polymerase. J Biol Chem. 1960 Aug;235:2399–2403. [PubMed] [Google Scholar]
  2. Banks G. R., Boezi J. A., Lehman I. R. A high molecular weight DNA polymerase from Drosophila melanogaster embryos. Purification, structure, and partial characterization. J Biol Chem. 1979 Oct 10;254(19):9886–9892. [PubMed] [Google Scholar]
  3. Banks G. R., Yarranton G. T. A DNA polymerase from Ustilago maydis. 2. Properties of the associated deoxyribonuclease activity. Eur J Biochem. 1976 Feb 2;62(1):143–150. doi: 10.1111/j.1432-1033.1976.tb10107.x. [DOI] [PubMed] [Google Scholar]
  4. Baril E. F., Brown O. E., Jenkins M. D., Laszlo J. Deoxyribonucleic acid polymerase with rat liver ribosomes and smooth membranes. Purification and properties of the enzymes. Biochemistry. 1971 May 25;10(11):1981–1992. doi: 10.1021/bi00787a004. [DOI] [PubMed] [Google Scholar]
  5. Baril E. F., Jenkins M. D., Brown O. E., Laszlo J., Morris H. P. DNA polymerases I and II in regenerating rat liver and Morris hepatomas. Cancer Res. 1973 Jun;33(6):1187–1193. [PubMed] [Google Scholar]
  6. Baril E., Mitchener J., Lee L., Baril B. Action of pancreatic DNase: requirements for activation of DNA as a template-primer for DNA polymerase. Nucleic Acids Res. 1977 Aug;4(8):2641–2653. doi: 10.1093/nar/4.8.2641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brakel C. L., Blumenthal A. B. Multiple forms of Drosophila embryo DNA polymerase: evidence for proteolytic conversion. Biochemistry. 1977 Jul 12;16(14):3137–3143. doi: 10.1021/bi00633a016. [DOI] [PubMed] [Google Scholar]
  8. Chang L. M., Bollum F. J. Low molecular weight deoxyribonucleic acid polymerase in mammalian cells. J Biol Chem. 1971 Sep 25;246(18):5835–5837. [PubMed] [Google Scholar]
  9. Chang L. M., Brown M., Bollum F. J. Induction of DNA polymerase in mouse L cells. J Mol Biol. 1973 Feb 15;74(1):1–8. doi: 10.1016/0022-2836(73)90349-5. [DOI] [PubMed] [Google Scholar]
  10. Chang L. M. DNA polymerases from bakers' yeast. J Biol Chem. 1977 Mar 25;252(6):1873–1880. [PubMed] [Google Scholar]
  11. Chang L. M. Low molecular weight deoxyribonucleic acid polymerase from calf thymus chromatin. I. Preparation of homogeneous enzyme. J Biol Chem. 1973 Jun 10;248(11):3789–3795. [PubMed] [Google Scholar]
  12. Chang L. M. Phylogeny of DNA polymerase-beta. Science. 1976 Mar 19;191(4232):1183–1185. doi: 10.1126/science.769158. [DOI] [PubMed] [Google Scholar]
  13. Chiu R. W., Baril E. F. Nuclear DNA polymerases and the HeLa cell cycle. J Biol Chem. 1975 Oct 10;250(19):7951–7957. [PubMed] [Google Scholar]
  14. Crerar M., Pearlman R. E. Deoxyribonucleic acid polymerase from Tetrahymena pyriformis. Purification and properties of the major activity in exponentially growing cells. J Biol Chem. 1974 May 25;249(10):3123–3131. [PubMed] [Google Scholar]
  15. Fridlender B., Fry M., Bolden A., Weissbach A. A new synthetic RNA-dependent DNA polymerase from human tissue culture cells (HeLa-fibroblast-synthetic oligonucleotides-template-purified enzymes). Proc Natl Acad Sci U S A. 1972 Feb;69(2):452–455. doi: 10.1073/pnas.69.2.452. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Furia M., Polito L. C., Locorotondo G., Grippo P. Identification of a DNA polymerase beta-like form in Drosophila melanogaster adult flies. Nucleic Acids Res. 1979 Jul 25;6(10):3399–3410. doi: 10.1093/nar/6.10.3399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Furukawa Y., Yamada R. H., Kohno M. Presence of two DNA polymerases in Tetrahymena pyriformis. Nucleic Acids Res. 1979 Dec 20;7(8):2387–2398. doi: 10.1093/nar/7.8.2387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Krokan H., Schaffer P., DePamphilis M. L. Involvement of eucaryotic deoxyribonucleic acid polymerases alpha and gamma in the replication of cellular and viral deoxyribonucleic acid. Biochemistry. 1979 Oct 2;18(20):4431–4443. doi: 10.1021/bi00587a025. [DOI] [PubMed] [Google Scholar]
  20. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  21. Longiaru M., Ikeda J. E., Jarkovsky Z., Horwitz S. B., Horwitz M. S. The effect of aphidicolin on adenovirus DNA synthesis. Nucleic Acids Res. 1979 Jul 25;6(10):3369–3386. doi: 10.1093/nar/6.10.3369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Loomis L. W., Rossomando E. F., Chang L. M. DNA polymerase of Dictyostelium discoideum. Biochim Biophys Acta. 1976 Apr 2;425(4):469–477. doi: 10.1016/0005-2787(76)90011-3. [DOI] [PubMed] [Google Scholar]
  23. McLennan A. G., Keir H. M. Deoxyribonucleic acid polymerases of Euglena gracilis. Purification and properties of two distinct deoxyribonucleic acid polymerases of high molecular weight. Biochem J. 1975 Nov;151(2):227–238. doi: 10.1042/bj1510227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Novak B., Baril E. F. HeLa DNA polymerase alpha activity in vitro: specific stimulation by a non-enzymic protein factor. Nucleic Acids Res. 1978 Jan;5(1):221–239. doi: 10.1093/nar/5.1.221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ohashi M., Taguchi T., Ikegami S. Aphidicolin: a specific inhibitor of DNA polymerases in the cytosol of rat liver. Biochem Biophys Res Commun. 1978 Jun 29;82(4):1084–1090. doi: 10.1016/0006-291x(78)90298-x. [DOI] [PubMed] [Google Scholar]
  26. Pederson T. Isolation and characterization of chromatin from the cellular slime mold, Dictyostelium discoideum. Biochemistry. 1977 Jun 14;16(12):2771–2777. doi: 10.1021/bi00631a029. [DOI] [PubMed] [Google Scholar]
  27. Spadari S., Weissbach A. The interrelation between DNA synthesis and various DNA polymerase activities in synchronized HeLa cells. J Mol Biol. 1974 Jun 15;86(1):11–20. doi: 10.1016/s0022-2836(74)80003-3. [DOI] [PubMed] [Google Scholar]
  28. Waqar M. A., Evans M. J., Huberman J. A. Effect of 2',3'-dideoxythymidine-5'-triphosphate on HeLa cell in vitro DNA synthesis: evidence that DNA polymerase alpha is the only polymerase required for cellular DNA replication. Nucleic Acids Res. 1978 Jun;5(6):1933–1946. doi: 10.1093/nar/5.6.1933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Weissbach A., Baltimore D., Bollum F., Gallo R., Korn D. Nomenclature of eukaryotic DNA polymerases. Science. 1975 Oct 24;190(4212):401–402. doi: 10.1126/science.1179222. [DOI] [PubMed] [Google Scholar]
  30. Weissbach A. Eukaryotic DNA polymerases. Annu Rev Biochem. 1977;46:25–47. doi: 10.1146/annurev.bi.46.070177.000325. [DOI] [PubMed] [Google Scholar]
  31. Weissbach A., Schlabach A., Fridlender B., Bolden A. DNA polymerases from human cells. Nat New Biol. 1971 Jun 9;231(23):167–170. doi: 10.1038/newbio231167a0. [DOI] [PubMed] [Google Scholar]
  32. Wintersberger U., Wintersberger E. DNA polymerases in polyoma virus-infected mouse kidney cells. J Virol. 1975 Nov;16(5):1095–1100. doi: 10.1128/jvi.16.5.1095-1100.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wintersberger U., Wintersberger E. Studies on deoxyribonucleic acid polymerases from yeast. 1. Parial purification and properties of two DNA polymerases from mitochondria-free cell extracts. Eur J Biochem. 1970 Mar 1;13(1):11–19. doi: 10.1111/j.1432-1033.1970.tb00893.x. [DOI] [PubMed] [Google Scholar]

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