Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 2000 Sep 15;350(Pt 3):757–763.

Novel triterpenoids inhibit both DNA polymerase and DNA topoisomerase.

Y Mizushina 1, A Iida 1, K Ohta 1, F Sugawara 1, K Sakaguchi 1
PMCID: PMC1221307  PMID: 10970789

Abstract

As described previously, we found that new triterpenoid compounds, designated fomitellic acids A and B, which selectively inhibit the activities of mammalian DNA polymerases alpha and beta [Mizushina, Tanaka, Kitamura, Tamai, Ikeda, Takemura, Sugawara, Arai, Matsukage, Yoshida and Sakaguchi (1998) Biochem. J. 330, 1325-1332; Tanaka, Kitamura, Mizushina, Sugawara and Sakaguchi (1998) J. Nat. Prod. 61, 193-197] and that a known triterpenoid, ursolic acid, is an inhibitor of human DNA topoisomerases I and II (A. Iida, Y. Mizushina and K. Sakaguchi, unpublished work). Here we report that all of these triterpenoids are potent inhibitors of calf DNA polymerase alpha, rat DNA polymerase beta and human DNA topoisomerases I and II, and show moderate inhibitory effects on plant DNA polymerase II and human immunodeficiency virus reverse transcriptase. However, these compounds did not influence the activities of prokaryotic DNA polymerases such as Escherichia coli DNA polymerase I or other DNA metabolic enzymes such as human telomerase, T7 RNA polymerase and bovine deoxyribonuclease I. These triterpenoids were not only mammalian DNA polymerase inhibitors but also inhibitors of DNA topoisomerases I and II even though the enzymic characteristics of DNA polymerases and DNA topoisomerases, including their modes of action, amino acid sequences and three-dimensional structures, differed markedly. These triterpenoids did not bind to DNA, suggesting that they act directly on these enzymes. Because the three-dimensional structures of fomitellic acids were shown by computer simulation to be very similar to that of ursolic acid, the DNA-binding sites of both enzymes, which compete for the inhibitors, might be very similar. Fomitellic acid A and ursolic acid prevented the growth of NUGC cancer cells, with LD(50) values of 38 and 30 microM respectively.

Full Text

The Full Text of this article is available as a PDF (210.0 KB).

Selected References

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

  1. Date T., Yamaguchi M., Hirose F., Nishimoto Y., Tanihara K., Matsukage A. Expression of active rat DNA polymerase beta in Escherichia coli. Biochemistry. 1988 Apr 19;27(8):2983–2990. doi: 10.1021/bi00408a048. [DOI] [PubMed] [Google Scholar]
  2. Ferro A. M., Olivera B. M. Poly(ADP-ribosylation) of DNA topoisomerase I from calf thymus. J Biol Chem. 1984 Jan 10;259(1):547–554. [PubMed] [Google Scholar]
  3. Kim N. W., Piatyszek M. A., Prowse K. R., Harley C. B., West M. D., Ho P. L., Coviello G. M., Wright W. E., Weinrich S. L., Shay J. W. Specific association of human telomerase activity with immortal cells and cancer. Science. 1994 Dec 23;266(5193):2011–2015. doi: 10.1126/science.7605428. [DOI] [PubMed] [Google Scholar]
  4. Kumar A., Abbotts J., Karawya E. M., Wilson S. H. Identification and properties of the catalytic domain of mammalian DNA polymerase beta. Biochemistry. 1990 Aug 7;29(31):7156–7159. doi: 10.1021/bi00483a002. [DOI] [PubMed] [Google Scholar]
  5. Kumar A., Widen S. G., Williams K. R., Kedar P., Karpel R. L., Wilson S. H. Studies of the domain structure of mammalian DNA polymerase beta. Identification of a discrete template binding domain. J Biol Chem. 1990 Feb 5;265(4):2124–2131. [PubMed] [Google Scholar]
  6. Liu L. F., Miller K. G. Eukaryotic DNA topoisomerases: two forms of type I DNA topoisomerases from HeLa cell nuclei. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3487–3491. doi: 10.1073/pnas.78.6.3487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Lu B. C., Sakaguchi K. An endo-exonuclease from meiotic tissues of the basidiomycete Coprinus cinereus. Its purification and characterization. J Biol Chem. 1991 Nov 5;266(31):21060–21066. [PubMed] [Google Scholar]
  8. Mizushina Y., Ohkubo T., Date T., Yamaguchi T., Saneyoshi M., Sugawara F., Sakaguchi K. Mode analysis of a fatty acid molecule binding to the N-terminal 8-kDa domain of DNA polymerase beta. A 1:1 complex and binding surface. J Biol Chem. 1999 Sep 3;274(36):25599–25607. doi: 10.1074/jbc.274.36.25599. [DOI] [PubMed] [Google Scholar]
  9. Mizushina Y., Takahashi N., Hanashima L., Koshino H., Esumi Y., Uzawa J., Sugawara F., Sakaguchi K. Lucidenic acid O and lactone, new terpene inhibitors of eukaryotic DNA polymerases from a basidiomycete, Ganoderma lucidum. Bioorg Med Chem. 1999 Sep;7(9):2047–2052. doi: 10.1016/s0968-0896(99)00121-2. [DOI] [PubMed] [Google Scholar]
  10. Mizushina Y., Tanaka N., Kitamura A., Tamai K., Ikeda M., Takemura M., Sugawara F., Arai T., Matsukage A., Yoshida S. The inhibitory effect of novel triterpenoid compounds, fomitellic acids, on DNA polymerase beta. Biochem J. 1998 Mar 15;330(Pt 3):1325–1332. doi: 10.1042/bj3301325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mizushina Y., Tanaka N., Yagi H., Kurosawa T., Onoue M., Seto H., Horie T., Aoyagi N., Yamaoka M., Matsukage A. Fatty acids selectively inhibit eukaryotic DNA polymerase activities in vitro. Biochim Biophys Acta. 1996 Sep 11;1308(3):256–262. doi: 10.1016/0167-4781(96)00121-2. [DOI] [PubMed] [Google Scholar]
  12. Mizushina Y., Watanabe I., Ohta K., Takemura M., Sahara H., Takahashi N., Gasa S., Sugawara F., Matsukage A., Yoshida S. Studies on inhibitors of mammalian DNA polymerase alpha and beta: sulfolipids from a pteridophyte, Athyrium niponicum. Biochem Pharmacol. 1998 Feb 15;55(4):537–541. doi: 10.1016/s0006-2952(97)00536-4. [DOI] [PubMed] [Google Scholar]
  13. Mizushina Y., Yagi H., Tanaka N., Kurosawa T., Seto H., Katsumi K., Onoue M., Ishida H., Iseki A., Nara T. Screening of inhibitor of eukaryotic DNA polymerases produced by microorganisms. J Antibiot (Tokyo) 1996 May;49(5):491–492. doi: 10.7164/antibiotics.49.491. [DOI] [PubMed] [Google Scholar]
  14. Mizushina Y., Yoshida S., Matsukage A., Sakaguchi K. The inhibitory action of fatty acids on DNA polymerase beta. Biochim Biophys Acta. 1997 Oct 20;1336(3):509–521. doi: 10.1016/s0304-4165(97)00067-6. [DOI] [PubMed] [Google Scholar]
  15. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983 Dec 16;65(1-2):55–63. doi: 10.1016/0022-1759(83)90303-4. [DOI] [PubMed] [Google Scholar]
  16. Murakami-Murofushi K., Kobayashi S., Onimura K., Matsumoto M., Shioda M., Yoshida S., Shoji M., Murofushi H. Selective inhibition of DNA polymerase-alpha family with chemically synthesized derivatives of PHYLPA, a unique Physarum lysophosphatidic acid. Biochim Biophys Acta. 1995 Aug 24;1258(1):57–60. doi: 10.1016/0005-2760(95)00097-v. [DOI] [PubMed] [Google Scholar]
  17. Murakami-Murofushi K., Shioda M., Kaji K., Yoshida S., Murofushi H. Inhibition of eukaryotic DNA polymerase alpha with a novel lysophosphatidic acid (PHYLPA) isolated from myxoamoebae of Physarum polycephalum. J Biol Chem. 1992 Oct 25;267(30):21512–21517. [PubMed] [Google Scholar]
  18. Nakayama C., Saneyoshi M. Inhibitory effects of 9-beta-D-xylofuranosyladenine 5'-triphosphate on DNA-dependent RNA polymerase I and II from cherry salmon (Oncorhynchus masou). J Biochem. 1985 May;97(5):1385–1389. doi: 10.1093/oxfordjournals.jbchem.a135192. [DOI] [PubMed] [Google Scholar]
  19. Ohta K., Mizushina Y., Hirata N., Takemura M., Sugawara F., Matsukage A., Yoshida S., Sakaguchi K. Action of a new mammalian DNA polymerase inhibitor, sulfoquinovosyldiacylglycerol. Biol Pharm Bull. 1999 Feb;22(2):111–116. doi: 10.1248/bpb.22.111. [DOI] [PubMed] [Google Scholar]
  20. Ohta K., Mizushina Y., Hirata N., Takemura M., Sugawara F., Matsukage A., Yoshida S., Sakaguchi K. Sulfoquinovosyldiacylglycerol, KM043, a new potent inhibitor of eukaryotic DNA polymerases and HIV-reverse transcriptase type 1 from a marine red alga, Gigartina tenella. Chem Pharm Bull (Tokyo) 1998 Apr;46(4):684–686. doi: 10.1248/cpb.46.684. [DOI] [PubMed] [Google Scholar]
  21. Ono K., Nakane H., Fukushima M. Differential inhibition of various deoxyribonucleic and ribonucleic acid polymerases by suramin. Eur J Biochem. 1988 Mar 1;172(2):349–353. doi: 10.1111/j.1432-1033.1988.tb13893.x. [DOI] [PubMed] [Google Scholar]
  22. Soltis D. A., Uhlenbeck O. C. Isolation and characterization of two mutant forms of T4 polynucleotide kinase. J Biol Chem. 1982 Oct 10;257(19):11332–11339. [PubMed] [Google Scholar]
  23. Sun N. J., Woo S. H., Cassady J. M., Snapka R. M. DNA polymerase and topoisomerase II inhibitors from Psoralea corylifolia. J Nat Prod. 1998 Mar;61(3):362–366. doi: 10.1021/np970488q. [DOI] [PubMed] [Google Scholar]
  24. Tamai K., Kojima K., Hanaichi T., Masaki S., Suzuki M., Umekawa H., Yoshida S. Structural study of immunoaffinity-purified DNA polymerase alpha-DNA primase complex from calf thymus. Biochim Biophys Acta. 1988 Sep 7;950(3):263–273. doi: 10.1016/0167-4781(88)90122-4. [DOI] [PubMed] [Google Scholar]
  25. Tanaka N., Kitamura A., Mizushina Y., Sugawara F., Sakaguchi K. Fomitellic acids, triterpenoid inhibitors of eukaryotic DNA polymerases from a basidiomycete, Fomitella fraxinea. J Nat Prod. 1998 Feb;61(2):193–197. doi: 10.1021/np970127a. [DOI] [PubMed] [Google Scholar]
  26. Ueno T., Takahashi H., Oda M., Mizunuma M., Yokoyama A., Goto Y., Mizushina Y., Sakaguchi K., Hayashi H. Inhibition of human telomerase by rubromycins: implication of spiroketal system of the compounds as an active moiety. Biochemistry. 2000 May 23;39(20):5995–6002. doi: 10.1021/bi992661i. [DOI] [PubMed] [Google Scholar]
  27. Wang J. C. DNA topoisomerases. Annu Rev Biochem. 1985;54:665–697. doi: 10.1146/annurev.bi.54.070185.003313. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES