Skip to main content
SpringerLink
Log in

Inhibitory action of conjugated C18-fatty acids on DNA polymerases and DNA topoisomerases

  • Articles
  • Published:
Lipids

Abstract

We reported previously that unsaturated linear-chain FA of the cis-configuration with a C18-hydrocarbon chain such as linoleic acid (18∶2Δ9c, 12c) could potently inhibit the activities of mammalian DNA polymerases and DNA topoisomerases, but their saturated forms could not. There are chemically two classes of unsaturated FA, normal and conjugated, but only the conjugated forms show potent antitumor activity. In this report, we study the inhibitory effects of chemically synthesized conjugated C18-FA on mammalian DNA polymerases and DNA topoisomerases as compared with normal unsaturated FA. The conjugated α-eleostearic acid (18∶3Δ9c, 11t, 13t) was the strongest of all the FA tested. For the inhibition, the conjugated form is crucially important. The energy-minimized 3-D structures of the FA were calculated, and both a length of less than 20 Å and a width of 8.13–9.24 Å in the C18-FA structure were found to be important for enzyme inhibition. The 3-D structure of the active site of both DNA polymerases and topoisomerases must have had a pocket to join α-eleostearic acid, and this pocket was 12.03 Å long and 9.24 Å wide.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

dsDNA:

double-stranded DNA

dTTP:

[3h]2′-deoxythymidine 5′-triphosphate

NP-40:

Nonidet P-40

oligo(dT)12–18 :

oligo(12–18) deoxyribothymidylic acid

pol:

DNA-directed DNA polymerase (EC 2.7.7.7)

poly(dA):

polydeoxyriboadenylic acid

poly(rC):

polycytidylic acid

topo:

DNA topoisomerase

References

  1. Mizushina, Y., Yagi, H., Tanaka, N., Kurosawa, T., Seto, H., Katumi, K., Onoue, M., Ishida, H., Iseki, A., Nara, T., et al. (1996) Screening of Inhibitor of Eukaryotic DNA Polymerases Produced by Microorganisms, J. Antibiot. (Tokyo) 49, 491–492.

    CAS  Google Scholar 

  2. Mizushina, Y., Tanaka, N., Yagi, H., Kurosawa, T., Onoue, M., Seto, H., Horie, T., Aoyagi, N., Yamaoka, M., Matsukage, A., et al. (1996) Fatty Acids Selectively Inhibit Eukaryotic DNA Polymerase Activities in vitro, Biochim. Biophys. Acta 1308, 256–262.

    PubMed  Google Scholar 

  3. Mizushina, Y., Yoshida, S., Matsukage, A., and Sakaguchi, K. (1997) The Inhibitory Action of Fatty Acids on DNA Polymerase β, Biochim. Biophys. Acta 1336, 509–521.

    PubMed  CAS  Google Scholar 

  4. Mizushina, Y., Ohkubo, T., Date, T., Yamaguchi, M., Saneyoshi, F., Sugawara, K., and Sakaguchi, K. (1999) Mode Analysis of a Fatty Acid Molecule Binding to the N-Terminal 8-kDa Domain of DNA Polymerase β, J. Biol. Chem. 274, 25599–25607.

    Article  PubMed  CAS  Google Scholar 

  5. Mizushina, Y., Sugawara, F., Iida, A., and Sakaguchi, K. (2000) Structural Homology Between DNA Binding Sites of DNA Polymerase β and DNA Topoisomerase II, J. Mol. Biol. 304, 385–395.

    Article  PubMed  CAS  Google Scholar 

  6. Ha, Y.L., Grimm, N.K., and Pariza, M.W. (1987) Anticarcinogens from Fried Ground Beef: Heat-Altered Derivatives of Linoleic Acid, Carcinogenesis 8, 1881–1887.

    PubMed  CAS  Google Scholar 

  7. Lee, K.N., Kritchevsky, D., and Pariza, M.W. (1994) Conjugated Linoleic Acid and Atherosclerosis in Rabbits, Atherosclerosis 108, 19–25.

    Article  PubMed  CAS  Google Scholar 

  8. Fritsche, J., and Steinhart, H. (1998) Analysis, Occurrence, and Physiological Properties of trans Fatty Acids (TFA) with Particular Emphasis on Conjugated Linoleic Acid Isomers (CLA)—A Review, Fett/Lipid 100, 190–210.

    Article  CAS  Google Scholar 

  9. Park, Y., Albright, K.J., Storkson, J.M., Liu, W., Cook, M.E., and Pariza, M.W. (1999) Changes in Body Composition in Mice During Feeding and Withdrawal of Conjugated Linoleic Acid, Lipids 34, 243–248.

    Article  PubMed  CAS  Google Scholar 

  10. Chin, S.F., Liu, W., Storkson, J.M., Ha, Y.L., and Pariza, M.W. (1992) Dietary Sources of Conjugated Dienoic Isomers of Linoleic Acid, a Newly Recognized Class of Anticarcinogens, J. Food Comp. Anal. 5, 185–197.

    Article  CAS  Google Scholar 

  11. Ip, C., Chin, S.F., Scimeca, J.A., and Pariza, M.W. (1991) Mammary Cancer Prevention by Conjugated Dienoic Derivative of Linoleic Acid, Cancer Res. 51, 6118–6124.

    PubMed  CAS  Google Scholar 

  12. Durgam, V.R., and Fernandes, G. (1997) The Growth Inhibitory Effect of Conjugated Linoleic Acid on MCF-7 Cells Is Related to Estrogen Response System, Cancer Lett. 116, 121–130.

    Article  PubMed  CAS  Google Scholar 

  13. Shultz, T.D., Chew, B.P., Seaman, W.R., and Luedecke, L.O. (1992) Inhibitory Effect of Conjugated Dienoic Derivatives of Linoleic Acid and β-Carotene on the in vitro Growth of Human Cancer Cells, Cancer Lett. 63, 125–133.

    Article  PubMed  CAS  Google Scholar 

  14. Schonberg, S., and Krokan, H.E. (1995) The Inhibitory Effect of Conjugated Dienoic Derivatives (CLA) of Linoleic Acid on the Growth of Human Tumor Cell Lines Is in Part Due to Increased Lipid Peroxidation, Anticancer Res. 15, 1241–1246.

    PubMed  CAS  Google Scholar 

  15. Cave, W.T., Jr. (1991) Dietary n−3 (omega-3) Polyunsaturated Fatty Acid Effects on Animal Tumorigenesis, FASEB J. 5, 2160–2166.

    PubMed  CAS  Google Scholar 

  16. Begin, M.E., Ells, G., and Horrobin, D.F. (1988) Polyunsaturated Fatty Acid-Induced Cytotoxicity Against Tumor Cells and Its Relationship to Lipid Peroxidation, J. Natl. Cancer Inst. 80, 188–194.

    PubMed  CAS  Google Scholar 

  17. Das, U.N. (1990) γ-Linolenic Acid, Arachidonic Acid, and Eicosapentaenoic Acid as Potential Anticancer Drugs, Nutrition 6, 429–434.

    PubMed  CAS  Google Scholar 

  18. Begin, M.E., Ells, G., Das, U.N., and Horrobin, D.F. (1986) Differential Killing of Human Carcinoma Cells Supplemented with n−3 and n−6 Polyunsaturated Fatty Acids, J. Natl. Cancer Inst. 77, 1053–1062.

    PubMed  CAS  Google Scholar 

  19. Tsuzuki, T., Igarashi, M., and Miyazawa, T. (2004) Conjugated Eicosapentanoic Acid Inhibits Transplanted Tumor Growth via Membrane Lipid Peroxidation in Nude Mice, J. Nutr. 134, 1162–1166.

    PubMed  CAS  Google Scholar 

  20. Tsuzuki, T., Tokuyama, Y., Igarashi, M., and Miyazawa, T. (2004) Tumor Growth Suppression by α-Eleostearic Acid, a Linolenic Acid Isomer with a Conjugated Triene System, via Lipid Peroxidation, Carcinogenesis 25, 1417–1425.

    Article  PubMed  CAS  Google Scholar 

  21. Weete, J.D. (1974) Fungal Lipid Biochemistry, Plenum Press, New York.

    Google Scholar 

  22. Association of Official Analytical Chemists (1990) Acids (polyunsaturated) in Oil and Fats, in Official Methods of Analysis of the Association of Official Analytical Chemists (Helrich, K., ed.), pp. 960–963, AOAC, Arlington, VA.

    Google Scholar 

  23. Pitt, G.A.J., and Morton, R.A. (1957) Ultra-violet Spectrophotometry of Fatty Acids, Prog. Chem. Fats Other Lipids 4, 227–278.

    Article  Google Scholar 

  24. Tamai, K., Kojima, K., Hanaichi, T., Masaki, S., Suzuki, M., Umekawa, H., and Yoshida, S. (1988) Structural Study of Immunoaffinity-Purified DNA Polymerase α-DNA Primase Complex from Calf Thymus, Biochim. Biophys. Acta 950, 263–273.

    PubMed  CAS  Google Scholar 

  25. Date, T., Yamaguchi, M., Hirose, F., Nishimoto, Y., Tanihara, K., and Matsukage, A. (1988) Expression of Active Rat DNA Polymerase β in Escherichia coli, Biochemistry 27, 2983–2990.

    Article  PubMed  CAS  Google Scholar 

  26. Oshige, M., Takeuchi, R., Ruike, R., Kuroda, K., and Sakaguchi, K. (2004) Subunit Protein-Affinity Isolation of Drosophila DNA Polymerase Catalytic Subunit, Protein Expr. Purif. 35, 248–256.

    Article  PubMed  CAS  Google Scholar 

  27. Suzuki, K., Shono, F., Kai, H., Uno, T., and Uyeda, M. (2000) Inhibition of Topoisomerases by Fatty Acids, J. Enzyme Inhib. 15, 357–366.

    Article  PubMed  CAS  Google Scholar 

  28. Sahara, H., Hanashima, S., Yamazaki, T., Takahashi, S., Sugawara, F., Ohtani, S., Ishikawa, M., Mizushina, Y., Ohta, K., Shimozawa, K., et al. (2002) Anti-tumor Effect of Chemically Synthesized Sulfolipids Based on Sea Urchin's Natural Sulfono-quinovosylmonoacylglycerols, Jpn. J. Cancer Res. 93, 85–92.

    PubMed  CAS  Google Scholar 

  29. Sakaguchi, K., Sugawara, F., and Mizushina, Y. (2002) Inhibitors of Eukaryotic DNA Polymerases, Seikagaku 74, 244–251.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratory of Food & Nutritional Sciences, Department of Nutritional Science, Kobe-Gakuin University, Nishi-ku, Kobe, 651-2180, Hyogo, Japan

    Yoshiyuki Mizushina, Kanako Kobayashi, Hiroshi Ikawa, Isoko Kuriyama, Yuko Yonezawa & Hiromi Yoshida

  2. High Technology Research Center, Kobe-Gakuin University, Nishi-ku, Kobe, 651-2180, Hyogo, Japan

    Yoshiyuki Mizushina & Hiromi Yoshida

  3. Food and Biodynamic Chemistry Laboratory, Graduate School of Life Science and Agriculture, Tohoku University, 981-8555, Sendai, Japan

    Tsuyoshi Tsuzuki, Takahiro Eitsuka & Teruo Miyazawa

  4. Life Science Research Center, Mie University, 514-8507, Tsu, Mie, Japan

    Masaharu Takemura

  5. Department of Applied Biological Science, Frontier Research Center for Genomic Drug Discovery, Tokyo University of Science, 278-8510, Noda-shi, Chiba, Japan

    Kengo Sakaguchi

Authors
  1. Yoshiyuki Mizushina
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tsuyoshi Tsuzuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Takahiro Eitsuka
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Teruo Miyazawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kanako Kobayashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hiroshi Ikawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Isoko Kuriyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yuko Yonezawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Masaharu Takemura
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Hiromi Yoshida
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Kengo Sakaguchi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yoshiyuki Mizushina.

About this article

Cite this article

Mizushina, Y., Tsuzuki, T., Eitsuka, T. et al. Inhibitory action of conjugated C18-fatty acids on DNA polymerases and DNA topoisomerases. Lipids 39, 977–983 (2004). https://doi.org/10.1007/s11745-004-1319-y

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11745-004-1319-y

Keywords

Search

Navigation