FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Biochemistry2009; 48(30); 7098-7109; doi: 10.1021/bi9006429

NMR and computational studies of stereoisomeric equine estrogen-derived DNA cytidine adducts in oligonucleotide duplexes: opposite orientations of diastereomeric forms.

Abstract: The equine estrogens equilin (EQ) and equilenin (EN) are the active components in the widely prescribed hormone replacement therapy formulation Premarin. Metabolic activation of EQ and EN generates the catechol 4-hydroxyequilenin (4-OHEN) that autoxidizes to the reactive o-quinone form in aerated aqueous solutions. The o-quinones react predominantly with C, and to a lesser extent with A and G, to form premutagenic cyclic covalent DNA adducts in vitro and in vivo. To obtain insights into the structural properties of these biologically important DNA lesions, we have synthesized site-specifically modified oligonucleotides containing the stereoisomeric 1'S,2'R,3'R-4-OHEN-C3 and 1'R,2'S,3'S-4-OHEN-C4 adducts derived from the reaction of 4-OHEN with the C in the oligonucleotide 5'-GGTAGCGATGG in aqueous solution. A combined NMR and computational approach was utilized to determine the conformational characteristics of the two major 4-OHEN-C3 and 4-OHEN-C4 stereoisomeric adducts formed in this oligonucleotide hybridized with its complementary strand. In both cases, the modified C adopts an anti glycosidic bond conformation; the equilenin distal ring protrudes into the minor groove while its two proximal hydroxyl groups are exposed on the major groove side of the DNA duplex. The bulky 4-OHEN-C adduct distorts the duplex within the central GC*G portion, but Watson-Crick pairing is maintained adjacent to C* in both stereoisomeric adducts. For the 4-OHEN-C3 adduct, the equilenin rings are oriented toward the 5'-end of the modified strand, while in 4-OHEN-C4 the equilenin is 3'-directed. Correspondingly, the distortions of the double-helical structures are more pronounced on the 5'- or the 3'-side of the lesion, respectively. These differences in stereoisomeric adduct conformations may play a role in the processing of these lesions in cellular environments.
Get Full Text
Publication Date: 2009-06-17 PubMed ID: 19527068PubMed Central: PMC2916639DOI: 10.1021/bi9006429Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article
  • Research Support
  • N.I.H.
  • Extramural

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research investigates the structural properties of DNA lesions obtained from equine estrogens equilin (EQ) and equilenin (EN), which are active components of a popular hormone therapy formulation. These estrogens can oxidize to form reactive compounds (o-quinones) that react with certain DNA components, creating pre-mutagenic DNA adducts. The study focused on understanding the structural impacts of these adducts on DNA, which can shed light on how they are processed within the cellular environment.

Study Objective and Methodology

  • This research aims to understand the structural properties of biologically significant DNA lesions resulting from equine estrogens EQ and EN. These estrogens are part of a commonly prescribed hormone replacement therapy and, when metabolically activated, can form compounds that interact with DNA, resulting in pre-mutagenic covalent DNA adducts.
  • The investigators synthesized specifically modified oligonucleotides, small DNA molecules, containing stereoisomeric 4-OHEN-C3 and 4-OHEN-C4 adducts. These adducts were generated from the reaction of 4-OHEN (a derivative of EQ and EN) with cytidine in an oligonucleotide sequence.
  • A combination of nuclear magnetic resonance (NMR) and computational studies was used to determine the conformational characteristics of these DNA adducts when the oligonucleotide combined with its complementary strand.

Findings and Interpretations

  • The study found that in both types of adducts, the modified cytidine adopts a specific conformation, with the equilenin ring protruding into the minor DNA groove and its two proximal hydroxyl groups being exposed on the major groove side of the DNA duplex.
  • These bulky adducts distort the DNA within the central part but maintain Watson-crick pairing next to the modified cytidine in both types of adducts.
  • The orientation of the equilenin rings and the resulting DNA distortions differ between the 4-OHEN-C3 and 4-OHEN-C4 adducts, with the distortions being more pronounced on the 5′- or the 3′- side of the lesion respectively.
  • These variations in the adduct conformations, observed from the study, can play an essential role in how these lesions are processed in cells and hence significantly impact the cellular environment and potentially cell function.

Significance of the Study

  • The research provides vital insights into the structural effects of equine estrogen-derived DNA adducts on the DNA structure which could further our understanding of its genetic impacts, especially in relation to its potential pre-mutagenic effects.
  • The findings from this study could be pivotal in understanding the potential carcinogenic risks associated with hormone replacement therapy and guide more targeted therapeutic strategies.

Cite This Article

APA
Zhang N, Ding S, Kolbanovskiy A, Shastry A, Kuzmin VA, Bolton JL, Patel DJ, Broyde S, Geacintov NE. (2009). NMR and computational studies of stereoisomeric equine estrogen-derived DNA cytidine adducts in oligonucleotide duplexes: opposite orientations of diastereomeric forms. Biochemistry, 48(30), 7098-7109. https://doi.org/10.1021/bi9006429

Publication

Biochemistry
ISSN: 1520-4995
NlmUniqueID: 0370623
Country: United States
Language: English
Volume: 48
Issue: 30
Pages: 7098-7109

Researcher Affiliations

Zhang, Na
  • Structural Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA.
Ding, Shuang
    Kolbanovskiy, Alexander
      Shastry, Anant
        Kuzmin, Vladimir A
          Bolton, Judy L
            Patel, Dinshaw J
              Broyde, Suse
                Geacintov, Nicholas E

                  MeSH Terms

                  • Animals
                  • Base Sequence
                  • Cytidine / chemistry
                  • Cytidine / metabolism
                  • DNA Adducts / chemistry
                  • DNA Damage
                  • Equilenin / analogs & derivatives
                  • Equilenin / chemistry
                  • Equilenin / metabolism
                  • Equilin / chemistry
                  • Equilin / metabolism
                  • Estradiol Congeners / chemistry
                  • Horses
                  • Humans
                  • Molecular Conformation
                  • Molecular Sequence Data
                  • Mutagenesis, Site-Directed
                  • Nuclear Magnetic Resonance, Biomolecular
                  • Nucleic Acid Conformation
                  • Oligonucleotides / chemistry
                  • Oligonucleotides / genetics
                  • Oligonucleotides / metabolism
                  • Stereoisomerism

                  Grant Funding

                  • CA-28038 / NCI NIH HHS
                  • R01 CA046533-20 / NCI NIH HHS
                  • R01 CA112412-05 / NCI NIH HHS
                  • R01 CA075449 / NCI NIH HHS
                  • R01 CA046533 / NCI NIH HHS
                  • CA-75449 / NCI NIH HHS
                  • R01 CA112412 / NCI NIH HHS
                  • CA-046533 / NCI NIH HHS
                  • R01 CA028038-29 / NCI NIH HHS
                  • R01 CA028038 / NCI NIH HHS
                  • R01 CA075449-12 / NCI NIH HHS
                  • CA112412 / NCI NIH HHS

                  References

                  This article includes 40 references
                  1. Hersh AL, Stefanick ML, Stafford RS. National use of postmenopausal hormone therapy: annual trends and response to recent evidence.. JAMA 2004 Jan 7;291(1):47-53.
                    pubmed: 14709575doi: 10.1001/jama.291.1.47google scholar: lookup
                  2. Rossouw JE, Anderson GL, Prentice RL, LaCroix AZ, Kooperberg C, Stefanick ML, Jackson RD, Beresford SA, Howard BV, Johnson KC, Kotchen JM, Ockene J. Risks and benefits of estrogen plus progestin in healthy postmenopausal women: principal results From the Women's Health Initiative randomized controlled trial.. JAMA 2002 Jul 17;288(3):321-33.
                    pubmed: 12117397doi: 10.1001/jama.288.3.321google scholar: lookup
                  3. Ravdin PM, Cronin KA, Howlader N, Berg CD, Chlebowski RT, Feuer EJ, Edwards BK, Berry DA. The decrease in breast-cancer incidence in 2003 in the United States.. N Engl J Med 2007 Apr 19;356(16):1670-4.
                    pubmed: 17442911doi: 10.1056/nejmsr070105google scholar: lookup
                  4. Yager JD, Davidson NE. Estrogen carcinogenesis in breast cancer.. N Engl J Med 2006 Jan 19;354(3):270-82.
                    pubmed: 16421368doi: 10.1056/nejmra050776google scholar: lookup
                  5. Okamoto Y, Chou PH, Kim SY, Suzuki N, Laxmi YR, Okamoto K, Liu X, Matsuda T, Shibutani S. Oxidative DNA damage in XPC-knockout and its wild mice treated with equine estrogen.. Chem Res Toxicol 2008 May;21(5):1120-4.
                    pubmed: 18447394doi: 10.1021/tx700428mgoogle scholar: lookup
                  6. Bolton JL. Quinoids, quinoid radicals, and phenoxyl radicals formed from estrogens and antiestrogens.. Toxicology 2002 Aug 1;177(1):55-65.
                    pubmed: 12126795doi: 10.1016/s0300-483x(02)00195-6google scholar: lookup
                  7. Zhang F, Chen Y, Pisha E, Shen L, Xiong Y, van Breemen RB, Bolton JL. The major metabolite of equilin, 4-hydroxyequilin, autoxidizes to an o-quinone which isomerizes to the potent cytotoxin 4-hydroxyequilenin-o-quinone.. Chem Res Toxicol 1999 Feb;12(2):204-13.
                    pubmed: 10027800doi: 10.1021/tx980217vgoogle scholar: lookup
                  8. Shen L, Qiu SX, van Breemen RB, Zhang FG, Chen YM, Bolton JL. Reaction of the Premarin metabolite 4-hydroxyequilenin semiquinone radical with 2′-deoxyguanosine: Formation of unusual cyclic adducts. Journal of the American Chemical Society 1997;119:11126–11127.
                  9. Shen L, Qiu S, Chen Y, Zhang F, van Breemen RB, Nikolic D, Bolton JL. Alkylation of 2'-deoxynucleosides and DNA by the Premarin metabolite 4-hydroxyequilenin semiquinone radical.. Chem Res Toxicol 1998 Feb;11(2):94-101.
                    pubmed: 9511900doi: 10.1021/tx970181rgoogle scholar: lookup
                  10. Embrechts J, Lemière F, Van Dongen W, Esmans EL. Equilenin-2'-deoxynucleoside adducts: analysis with nano-liquid chromatography coupled to nano-electrospray tandem mass spectrometry.. J Mass Spectrom 2001 Mar;36(3):317-28.
                    pubmed: 11312524doi: 10.1002/jms.136google scholar: lookup
                  11. Kolbanovskiy A, Kuzmin V, Shastry A, Kolbanovskaya M, Chen D, Chang M, Bolton JL, Geacintov NE. Base selectivity and effects of sequence and DNA secondary structure on the formation of covalent adducts derived from the equine estrogen metabolite 4-hydroxyequilenin.. Chem Res Toxicol 2005 Nov;18(11):1737-47.
                    pubmed: 16300383doi: 10.1021/tx050190xgoogle scholar: lookup
                  12. Bolton JL, Thatcher GR. Potential mechanisms of estrogen quinone carcinogenesis.. Chem Res Toxicol 2008 Jan;21(1):93-101.
                    pmc: PMC2556295pubmed: 18052105doi: 10.1021/tx700191pgoogle scholar: lookup
                  13. Zhang F, Swanson SM, van Breemen RB, Liu X, Yang Y, Gu C, Bolton JL. Equine estrogen metabolite 4-hydroxyequilenin induces DNA damage in the rat mammary tissues: formation of single-strand breaks, apurinic sites, stable adducts, and oxidized bases.. Chem Res Toxicol 2001 Dec;14(12):1654-9.
                    pubmed: 11743748doi: 10.1021/tx010158cgoogle scholar: lookup
                  14. Embrechts J, Lemière F, Van Dongen W, Esmans EL, Buytaert P, Van Marck E, Kockx M, Makar A. Detection of estrogen DNA-adducts in human breast tumor tissue and healthy tissue by combined nano LC-nano ES tandem mass spectrometry.. J Am Soc Mass Spectrom 2003 May;14(5):482-91.
                    pubmed: 12745217doi: 10.1016/s1044-0305(03)00130-2google scholar: lookup
                  15. Suzuki N, Yasui M, Santosh Laxmi YR, Ohmori H, Hanaoka F, Shibutani S. Translesion synthesis past equine estrogen-derived 2'-deoxycytidine DNA adducts by human DNA polymerases eta and kappa.. Biochemistry 2004 Sep 7;43(35):11312-20.
                    pubmed: 15366941doi: 10.1021/bi049273ngoogle scholar: lookup
                  16. Yasui M, Laxmi YR, Ananthoju SR, Suzuki N, Kim SY, Shibutani S. Translesion synthesis past equine estrogen-derived 2'-deoxyadenosine DNA adducts by human DNA polymerases eta and kappa.. Biochemistry 2006 May 16;45(19):6187-94.
                    pmc: PMC2504361pubmed: 16681391doi: 10.1021/bi0525324google scholar: lookup
                  17. Ding S, Wang Y, Kolbanovskiy A, Durandin A, Bolton JL, van Breemen RB, Broyde S, Geacintov NE. Determination of absolute configurations of 4-hydroxyequilenin-cytosine and -adenine adducts by optical rotatory dispersion, electronic circular dichroism, density functional theory calculations, and mass spectrometry.. Chem Res Toxicol 2008 Sep;21(9):1739-48.
                    pmc: PMC2574695pubmed: 18680315doi: 10.1021/tx800095fgoogle scholar: lookup
                  18. Ding S, Shapiro R, Geacintov NE, Broyde S. Conformations of stereoisomeric base adducts to 4-hydroxyequilenin.. Chem Res Toxicol 2003 Jun;16(6):695-707.
                    pubmed: 12807352doi: 10.1021/tx0340246google scholar: lookup
                  19. Ding S, Shapiro R, Geacintov NE, Broyde S. Equilenin-derived DNA adducts to cytosine in DNA duplexes: structures and thermodynamics.. Biochemistry 2005 Nov 8;44(44):14565-76.
                    pubmed: 16262256doi: 10.1021/bi051090tgoogle scholar: lookup
                  20. Ding S, Shapiro R, Geacintov NE, Broyde S. 4-hydroxyequilenin-adenine lesions in DNA duplexes: stereochemistry, damage site, and structure.. Biochemistry 2007 Jan 9;46(1):182-91.
                    pmc: PMC2582198pubmed: 17198388doi: 10.1021/bi061652ogoogle scholar: lookup
                  21. Ding S, Shapiro R, Cai Y, Geacintov NE, Broyde S. Conformational properties of equilenin-DNA adducts: stereoisomer and base effects.. Chem Res Toxicol 2008 May;21(5):1064-73.
                    pmc: PMC2597795pubmed: 18416538doi: 10.1021/tx800010ugoogle scholar: lookup
                  22. Chen D. Nucleotide Excision Repair and Translesion Synthesis of DNA Adducts Derived from the Equine Estrogen Metabolite 4-hydroxyequilenin. Department of Chemistry. New York University; New York: 2006.
                  23. Chen DD, Oum L, Kolbanovskiy A, Kuzmin V, Shastry A, Chang MS, Bolton JL, Geacintov N. Translesion synthesis and nucleotide excision repair of site specifically modified oligodeoxyribonucleotides containing single lesions derived from the equine estrogen metabolite 4-OHEN. Chem Res Toxicol 2004;17:1782–1782.
                  24. Cai Y, Patel DJ, Geacintov NE, Broyde S. Differential nucleotide excision repair susceptibility of bulky DNA adducts in different sequence contexts: hierarchies of recognition signals.. J Mol Biol 2009 Jan 9;385(1):30-44.
                    pmc: PMC2680230pubmed: 18948114doi: 10.1016/j.jmb.2008.09.087google scholar: lookup
                  25. Geacintov NE, Broyde S, Buterin T, Naegeli H, Wu M, Yan S, Patel DJ. Thermodynamic and structural factors in the removal of bulky DNA adducts by the nucleotide excision repair machinery.. Biopolymers 2002 Nov 5;65(3):202-10.
                    pubmed: 12228925doi: 10.1002/bip.10239google scholar: lookup
                  26. Case DA, Darden TA, Cheatham TE III, Simmerling CL, Wang J, Duke RE, Luo R, Merz KM, Wang B, Pearlman DA, Crowley M, Brozell S, Tsui V, Gohlke H, Mongan J, Hornak V, Cui G, Beroza P, Schafmeister C, Kollman PA. AMBER 8. University of California; San Francisco, CA: 2004.
                  27. Srinivasan J, Cheatham TE, Cieplak P, Kollman PA, Case DA. Continuum solvent studies of the stability of DNA, RNA, and phosphoramidate - DNA helices. Journal of the American Chemical Society 1998;120:9401–9409.
                  28. Kollman PA, Massova I, Reyes C, Kuhn B, Huo S, Chong L, Lee M, Lee T, Duan Y, Wang W, Donini O, Cieplak P, Srinivasan J, Case DA, Cheatham TE 3rd. Calculating structures and free energies of complex molecules: combining molecular mechanics and continuum models.. Acc Chem Res 2000 Dec;33(12):889-97.
                    pubmed: 11123888doi: 10.1021/ar000033jgoogle scholar: lookup
                  29. Simmerling C, Elber R, Zhang J. MOIL-View—a program for visualization of structure and dynamics of biomolecules and STO—a program for computing stochastic paths. In: Pullman A, editor. Modeling of Biomolecular Structure and Mechanisms. Kluwer; Netherlands: 1995.
                  30. Hingerty BE, Figueroa S, Hayden TL, Broyde S. Prediction of DNA structure from sequence: a build-up technique.. Biopolymers 1989 Jul;28(7):1195-222.
                    pubmed: 2775836doi: 10.1002/bip.360280703google scholar: lookup
                  31. Ravishanker G, Swaminathan S, Beveridge DL, Lavery R, Sklenar H. Conformational and helicoidal analysis of 30 PS of molecular dynamics on the d(CGCGAATTCGCG) double helix: "curves", dials and windows.. J Biomol Struct Dyn 1989 Feb;6(4):669-99.
                    pubmed: 2619934doi: 10.1080/07391102.1989.10507729google scholar: lookup
                  32. Ravishanker G, Wang W, Beveridge DL. MD Toolchest. Wesleyan University; Middletown, CT:
                  33. Patel DJ, Kozlowski SA, Nordheim A, Rich A. Right-handed and left-handed DNA: studies of B- and Z-DNA by using proton nuclear Overhauser effect and P NMR.. Proc Natl Acad Sci U S A 1982 Mar;79(5):1413-7.
                    pmc: PMC345983pubmed: 6951185doi: 10.1073/pnas.79.5.1413google scholar: lookup
                  34. Patel DJ, Shapiro L, Hare D. DNA and RNA: NMR studies of conformations and dynamics in solution.. Q Rev Biophys 1987 Aug;20(1-2):35-112.
                    pubmed: 2448843doi: 10.1017/s0033583500004224google scholar: lookup
                  35. Van de Ven FJ, Hilbers CW. Nucleic acids and nuclear magnetic resonance.. Eur J Biochem 1988 Dec 1;178(1):1-38.
                    pubmed: 3060357doi: 10.1111/j.1432-1033.1988.tb14425.xgoogle scholar: lookup
                  36. Kouchakdjian M, Marinelli E, Gao XL, Johnson F, Grollman A, Patel D. NMR studies of exocyclic 1,N2-propanodeoxyguanosine adducts (X) opposite purines in DNA duplexes: protonated X(syn).A(anti) pairing (acidic pH) and X(syn).G(anti) pairing (neutral pH) at the lesion site.. Biochemistry 1989 Jun 27;28(13):5647-57.
                    pubmed: 2775729doi: 10.1021/bi00439a047google scholar: lookup
                  37. de los Santos C, Kouchakdjian M, Yarema K, Basu A, Essigmann J, Patel DJ. NMR studies of the exocyclic 1,N6-ethenodeoxyadenosine adduct (epsilon dA) opposite deoxyguanosine in a DNA duplex. Epsilon dA(syn).dG(anti) pairing at the lesion site.. Biochemistry 1991 Feb 19;30(7):1828-35.
                    pubmed: 1993197doi: 10.1021/bi00221a015google scholar: lookup
                  38. Yasui M, Matsui S, Laxmi YR, Suzuki N, Kim SY, Shibutani S, Matsuda T. Mutagenic events induced by 4-hydroxyequilin in supF shuttle vector plasmid propagated in human cells.. Carcinogenesis 2003 May;24(5):911-7.
                    pubmed: 12771036doi: 10.1093/carcin/bgg029google scholar: lookup
                  39. Yasui M, Suzuki N, Liu X, Okamoto Y, Kim SY, Laxmi YR, Shibutani S. Mechanism of translesion synthesis past an equine estrogen-DNA adduct by Y-family DNA polymerases.. J Mol Biol 2007 Aug 31;371(5):1151-62.
                    pmc: PMC2039719pubmed: 17603077doi: 10.1016/j.jmb.2007.06.009google scholar: lookup
                  40. Lu XJ, Olson WK. 3DNA: a software package for the analysis, rebuilding and visualization of three-dimensional nucleic acid structures.. Nucleic Acids Res 2003 Sep 1;31(17):5108-21.
                    pmc: PMC212791pubmed: 12930962doi: 10.1093/nar/gkg680google scholar: lookup

                  Citations

                  This article has been cited 4 times.
                  1. Liu Y, Reeves D, Kropachev K, Cai Y, Ding S, Kolbanovskiy M, Kolbanovskiy A, Bolton JL, Broyde S, Van Houten B, Geacintov NE. Probing for DNA damage with β-hairpins: similarities in incision efficiencies of bulky DNA adducts by prokaryotic and human nucleotide excision repair systems in vitro. DNA Repair (Amst) 2011 Jul 15;10(7):684-96.
                    doi: 10.1016/j.dnarep.2011.04.020pubmed: 21741328google scholar: lookup
                  2. Okahashi Y, Iwamoto T, Suzuki N, Shibutani S, Sugiura S, Itoh S, Nishiwaki T, Ueno S, Mori T. Quantitative detection of 4-hydroxyequilenin-DNA adducts in mammalian cells using an immunoassay with a novel monoclonal antibody. Nucleic Acids Res 2010 Jul;38(12):e133.
                    doi: 10.1093/nar/gkq233pubmed: 20406772google scholar: lookup
                  3. Noschang CG, Krolow R, Fontella FU, Arcego DM, Diehl LA, Weis SN, Arteni NS, Dalmaz C. Neonatal handling impairs spatial memory and leads to altered nitric oxide production and DNA breaks in a sex specific manner. Neurochem Res 2010 Jul;35(7):1083-91.
                    doi: 10.1007/s11064-010-0158-7pubmed: 20369293google scholar: lookup
                  4. Ding S, Kolbanovskiy A, Durandin A, Crean C, Shafirovich V, Broyde S, Geacintov NE. Absolute configurations of DNA lesions determined by comparisons of experimental ECD and ORD spectra with DFT calculations. Chirality 2009;21 Suppl 1(0 1):E231-41.
                    doi: 10.1002/chir.20804pubmed: 19937959google scholar: lookup
                  Subscribe to our newsletter
                  Join 99,780 horse owners like you who receive our email updates on horse health and nutrition.