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Home / Equine Research Bank / Biochemistry / NMR and computational studies of stereoisomeric equine estro...
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.
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Publication Date: 2009-06-17 PubMed ID: 19527068PubMed Central: PMC2916639DOI: 10.1021/bi9006429Google Scholar: Lookup
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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

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                  Citations

                  This article has been cited 4 times.
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