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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
. 1984 May;81(9):2635–2639. doi: 10.1073/pnas.81.9.2635

Hydrolysis of benzo[a]pyrene diol epoxide and its covalent binding to DNA proceed through similar rate-determining steps.

T Meehan, D M Bond
PMCID: PMC345124  PMID: 6425834

Abstract

The mutagenic and carcinogenic metabolite of benzo[a]pyrene, (7R,8S)-dihydroxy-(9R,10R)-epoxy-7,8, 9,10-tetrahydrobenzo[a]pyrene, undergoes two major reactions in the presence of DNA: (i) hydrolysis and (ii) covalent binding. We report that hydrolysis and covalent binding are specific and general acid-catalyzed reactions with the same or similar rate-determining steps. To account for the similarity of rate-determining steps in covalent binding and hydrolysis we propose and test two models. In each model, the rate-determining step results in formation of a carbonium ion, which serves as a precursor for both tetrol and adduct. In model A the carbonium ion is partitioned between two domains (1 and 2), while in model B there is only one domain. Measurements of pseudo-first-order rate constants, product ratios, and rate ratios support model A, while kinetic results are inconsistent with model B. Domain 1 most likely represents activated benzo[a]pyrenes that are intercalated into DNA, while domain 2 hydrocarbons are physically bound to the outside of the DNA helix.

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Selected References

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

  1. Drinkwater N. R., Miller J. A., Miller E. C., Yang N. C. Covalent intercalative binding to DNA in relation to the mutagenicity of hydrocarbon epoxides and N-acetoxy-2-acetylaminofluorene. Cancer Res. 1978 Oct;38(10):3247–3255. [PubMed] [Google Scholar]
  2. Gamper H. B., Straub K., Calvin M., Bartholomew J. C. DNA alkylation and unwinding induced by benzo[a]pyrene diol epoxide: modulation by ionic strength and superhelicity. Proc Natl Acad Sci U S A. 1980 Apr;77(4):2000–2004. doi: 10.1073/pnas.77.4.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Geacintov N. E., Gagliano A., Ivanovic V., Weinstein I. B. Electric linear dichroism study on the orientation of benzo[alpha]pyrene-7,8-dihydrodiol 9,10-oxide covalently bound to DNA. Biochemistry. 1978 Nov 28;17(24):5256–5262. doi: 10.1021/bi00617a027. [DOI] [PubMed] [Google Scholar]
  4. Geacintov N. E., Ibanez V., Gagliano A. G., Yoshida H., Harvey R. G. Kinetics of hydrolysis to tetraols and binding of benzo(a)pyrene-7,8-dihydrodiol-9, 10-oxide and its tetraol derivatives to DNA. Conformation of adducts. Biochem Biophys Res Commun. 1980 Feb 27;92(4):1335–1342. doi: 10.1016/0006-291x(80)90432-5. [DOI] [PubMed] [Google Scholar]
  5. Geacintov N. E., Yoshida H., Ibanez V., Harvey R. G. Non-covalent intercalative binding of 7,8-dihydroxy-9,10-epoxybenzo(a)pyrene to DNA. Biochem Biophys Res Commun. 1981 Jun;100(4):1569–1577. doi: 10.1016/0006-291x(81)90698-7. [DOI] [PubMed] [Google Scholar]
  6. Hogan M. E., Dattagupta N., Whitlock J. P., Jr Carcinogen-induced alteration of DNA structure. J Biol Chem. 1981 May 10;256(9):4504–4513. [PubMed] [Google Scholar]
  7. Huberman E., Sachs L., Yang S. K., Gelboin V. Identification of mutagenic metabolites of benzo(a)pyrene in mammalian cells. Proc Natl Acad Sci U S A. 1976 Feb;73(2):607–611. doi: 10.1073/pnas.73.2.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ivanovic V., Geacintov N. E., Yamasaki H., Weinstein I. B. DNA and RNA adducts formed in hamster embryo cell cultures exposed to benzo[a]pyrene. Biochemistry. 1978 May 2;17(9):1597–1603. doi: 10.1021/bi00602a003. [DOI] [PubMed] [Google Scholar]
  9. Jeffrey A. M., Grzeskowiak K., Weinstein I. B., Nakanishi K., Roller P., Harvey R. G. Benzo(a)pyrene-7,8-dihydrodiol 9,10-oxide adenosine and deoxyadenosine adducts: structure and stereochemistry. Science. 1979 Dec 14;206(4424):1309–1311. doi: 10.1126/science.316186. [DOI] [PubMed] [Google Scholar]
  10. King H. W., Osborne M. R., Beland F. A., Harvey R. G., Brookes P. (+/-)-7alpha,8beta-dihydroxy-9beta,10beta-epoxy-7,8,9,10-tetrahydrobenzo(a)-pyrene is an intermediate in the metabolism and binding to DNA of benzo(a)pyrene. Proc Natl Acad Sci U S A. 1976 Aug;73(8):2679–2681. doi: 10.1073/pnas.73.8.2679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kootstra A., Haas B. L., Slaga T. J. Reactions of benzo[a]pyrene diol-epoxides with DNA and nucleosomes in aqueous solutions. Biochem Biophys Res Commun. 1980 Jun 30;94(4):1432–1438. doi: 10.1016/0006-291x(80)90579-3. [DOI] [PubMed] [Google Scholar]
  12. Koreeda M., Moore P. D., Yagi H., Yeh H. J., Jerina D. M. Alkylation of polyguanylic acid at the 2-amino group and phosphate by the potent mutagen (+/-)-7beta,8alpha-dihydroxy-9beta,10beta-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene. J Am Chem Soc. 1976 Oct 13;98(21):6720–6722. doi: 10.1021/ja00437a061. [DOI] [PubMed] [Google Scholar]
  13. Li H. J., Crothers D. M. Relaxation studies of the proflavine-DNA complex: the kinetics of an intercalation reaction. J Mol Biol. 1969 Feb 14;39(3):461–477. doi: 10.1016/0022-2836(69)90138-7. [DOI] [PubMed] [Google Scholar]
  14. McCann J., Ames B. N. Detection of carcinogens as mutagens in the Salmonella/microsome test: assay of 300 chemicals: discussion. Proc Natl Acad Sci U S A. 1976 Mar;73(3):950–954. doi: 10.1073/pnas.73.3.950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Meehan T., Gamper H., Becker J. F. Characterization of reversible, physical binding of benzo[a]pyrene derivatives to DNA. J Biol Chem. 1982 Sep 10;257(17):10479–10485. [PubMed] [Google Scholar]
  16. Meehan T., Straub K., Calvin M. Benzo[alpha]pyrene diol epoxide covalently binds to deoxyguanosine and deoxyadenosine in DNA. Nature. 1977 Oct 20;269(5630):725–727. doi: 10.1038/269725a0. [DOI] [PubMed] [Google Scholar]
  17. Meehan T., Straub K., Calvin M. Elucidation of hydrocarbon structure in an enzyme-catalyzed benzo[a]pyrene-poly (G) covalent complex. Proc Natl Acad Sci U S A. 1976 May;73(5):1437–1441. doi: 10.1073/pnas.73.5.1437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Meehan T., Straub K. Double-stranded DNA steroselectively binds benzo(a)pyrene diol epoxides. Nature. 1979 Feb 1;277(5695):410–412. doi: 10.1038/277410a0. [DOI] [PubMed] [Google Scholar]
  19. Michaud D. P., Gupta S. C., Whalen D. L., Sayer J. M., Jerina D. M. Effects of pH and salt concentration on the hydrolysis of a benzo[alpha]pyrene 7,8-diol-9,10-epoxide catalyzed by DNA and polyadenylic acid. Chem Biol Interact. 1983 Apr-May;44(1-2):41–52. doi: 10.1016/0009-2797(83)90128-x. [DOI] [PubMed] [Google Scholar]
  20. Osborne M. R., Harvey R. G., Brookes P. The reaction of trans-7,8-dihydroxy-anti-9,10-epoxy-7,8,9,10-tetrahydrobenzo(a)pyrene with DNA involves attack at the N7-position of guanine moieties. Chem Biol Interact. 1978 Jan;20(1):123–130. doi: 10.1016/0009-2797(78)90087-x. [DOI] [PubMed] [Google Scholar]
  21. Sims P., Grover P. L., Swaisland A., Pal K., Hewer A. Metabolic activation of benzo(a)pyrene proceeds by a diol-epoxide. Nature. 1974 Nov 22;252(5481):326–328. doi: 10.1038/252326a0. [DOI] [PubMed] [Google Scholar]
  22. Straub K. M., Meehan T., Burlingame A. L., Calvin M. Identification of the major adducts formed by reaction of benzo(a)pyrene diol epoxide with DNA in vitro. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5285–5289. doi: 10.1073/pnas.74.12.5285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Weinstein I. B., Jeffrey A. M., Jennette K. W., Blobstein S. H., Harvey R. G., Harris C., Autrup H., Kasai H., Nakanishi K. Benzo(a)pyrene diol epoxides as intermediates in nucleic acid binding in vitro and in vivo. Science. 1976 Aug 13;193(4253):592–595. doi: 10.1126/science.959820. [DOI] [PubMed] [Google Scholar]
  24. Wood A. W., Sayer J. M., Newmark H. L., Yagi H., Michaud D. P., Jerina D. M., Conney A. H. Mechanism of the inhibition of mutagenicity of a benzo[a]pyrene 7,8-diol 9,10-epoxide by riboflavin 5'-phosphate. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5122–5126. doi: 10.1073/pnas.79.17.5122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Yang S. K., McCourt D. W., Gelboin H. V. The mechanism of hydrolysis of the non-K-region benzo[a]pyrene diol epoxide r-7, t-8-dihydroxy-t-9,10-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene. J Am Chem Soc. 1977 Jul 20;99(15):5130–5134. doi: 10.1021/ja00457a037. [DOI] [PubMed] [Google Scholar]

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