DNA sequence‐specific adducts of adriamycin and mitomycin C

Phillips, Don R.; White, Robin J.; Cullinane, Carleen

doi:10.1016/0014-5793(89)80289-3

Cited by 44 publications

(40 citation statements)

References 25 publications

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“…Our ability to produce and purify large amounts of stable covalent Adriamycin-DNA adduct in solution has allowed a more direct structural comparison of these adducts than previously possible by MS. CH 2 O-and Tris͞Fe(III)͞DTT-mediated Adriamycin-DNA adducts were formed on the selfcomplementary hexanucleotide d(ATGCAT) 2 , containing only a single potential site for adduct formation. Isolation and purification of the respective adduct and analysis by high-field 1 H NMR indicated that the CH 2 O-and Tris͞Fe(III)͞DTT-mediated Adriamycin-DNA adducts are structurally identical (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Fig. 3 shows the anomeric-aromatic portion of the two-dimensional 1 H NOESY spectrum of the Adriamycin-DNA adduct formed on d(ATGCAT) 2 . From the connectivity pattern in this region, it is clear that only one reaction product is present, in which a single Adriamycin molecule is covalently bound per DNA duplex.…”

Section: Incorporation Of 13 C -Labeled Ch 2 Omentioning

confidence: 99%

“…Some years ago, in vitro transcription analysis indicated that Adriamycin can form covalent adducts with DNA (2). These occurred almost exclusively at GpC sequences and were found to behave much as classical interstrand DNA cross-links in their ability to block RNA polymerase (3,4).…”

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confidence: 99%

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Characterization of covalent Adriamycin-DNA adducts

Zeman

Phillips

Crothers

1998

Proc. Natl. Acad. Sci. U.S.A.

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146

142

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Adriamycin is a popular antineoplastic agent whose ability to form covalent adducts with DNA has been correlated to cellular apoptosis (programmed cell death) in tumor models. We have isolated and purified this adduct formed under oxido-reductive (Fenton) conditions in Tris buffer. We show by homo-and heteronuclear NMR spectroscopy that the covalent Adriamycin-DNA adduct is structurally equivalent to that resulting from direct reaction with formaldehyde. Covalent linkage of the drug to one of the DNA strands confers remarkable stability to the duplex, indicated by a 162-fold reduction in the rate of strand displacement compared with the complex with noncovalently bound drug. Glyceraldehyde also engenders covalent Adriamycin-DNA complexes, providing a possible relevant biological context for in vivo adduct formation.

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Section: Resultsmentioning

confidence: 99%

Section: Incorporation Of 13 C -Labeled Ch 2 Omentioning

confidence: 99%

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Characterization of covalent Adriamycin-DNA adducts

Zeman

Phillips

Crothers

1998

Proc. Natl. Acad. Sci. U.S.A.

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146

142

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“…[100] Phillips, White, and Cullinane implemented an in vitro transcription assay exhibiting blockage of transcription by E. coli RNA polymerase at specific DNA sites when mitomycin C was reduced by xanthine oxidase/NADH, that showed selectivity for XpC sequences of the non-coding strand, corresponding to G selectivity of the coding strand. [101] Through experiments in the Hopkins lab involving sequence-random cleavage of MMC cross-linked DNA, the sequence specificity of the cross links was determined at single nucleotide resolution to be at the G of 5′- d(CG) in strong preference to 5′-d(GC). [102] Crothers simultaneously utilized a gel electrophoresis assay yielding identical results.…”

Section: Biochemistrymentioning

confidence: 99%

Mitomycinoid Alkaloids: Mechanism of Action, Biosynthesis, Total Syntheses, and Synthetic Approaches

et al. 2013

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“…1) that lies in the minor groove without forming any bonds to DNA (4), although eventually it may also form covalent adducts with DNA (36,37).…”

Section: Discussionmentioning

confidence: 99%

Daunomycin-induced Unfolding and Aggregation of Chromatin

Rabbani

Maya

Ausió

1999

Journal of Biological Chemistry

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Using equilibrium dialysis and sedimentation velocity analysis, we have characterized the binding of the antitumor drug daunomycin to chicken erythrocyte chromatin before and after depletion of linker histones and to its constitutive DNA under several ionic strengths (5, 25, and 75 mM NaCl). The equilibrium dialysis experiments reveal that the drug binds cooperatively to both the chromatin fractions and to the DNA counterpart within the range of ionic strength used in this study. A significant decrease in the binding affinity was observed at 75 mM NaCl. At any given salt concentration, daunomycin exhibits higher binding affinity for DNA than for linker histone-depleted chromatin or chromatin (in decreasing order). Binding of daunomycin to DNA does not significantly affect the sedimentation coefficient of the molecule. This is in contrast to binding to chromatin and to its linker histone-depleted counterpart. In these instances, preferential binding of the drug to the linker DNA regions induces an unfolding of the chromatin fiber that is followed by aggregation, presumably because of histone-DNA interfiber interactions.Daunomycin is an anthracycline antibiotic widely used as a potent chemotherapeutic agent in the treatment of various cancers (1, 2). Studies on its mechanism of action indicate that nuclear DNA is an important target for daunomycin. The structure of this pharmacologically active drug consists of two distinct domains ( Fig. 1): a planar aglycon chromophore that intercalates between adjacent base pairs of DNA and an amino sugar ring that lies in the minor groove of the DNA double helix (3, 4). Binding of daunomycin to DNA results in the inhibition of both DNA replication and RNA transcription (5-7).The binding of daunomycin to DNA has been studied in detail in the past (8 -11). However, in the cell, DNA does not exist as a naked structure but is associated with histones and other nuclear proteins in a complex that is known as chromatin (12-13). Thus, chromatin, not DNA, is the major target for daunomycin in vivo. How the presence of chromosomal proteins might modulate the binding of this drug to DNA and how in turn chromatin structure is affected by the binding of the drug are important questions that need to answered to understand the biological mechanism of action of daunomycin. Although several attempts in this direction have already been carried out using isolated nucleosome (14, 15) and crude chromatin fractions (16 -18), the mechanism of daunomycin binding to chromatin and the conformational transitions involved are still poorly understood. In this paper we report several experiments designed to address these questions in more detail.We have studied the ionic strength dependence of the binding of daunomycin to a well defined fraction of chicken erythrocyte chromatin consisting of an average number of 43 nucleosomes. The results are compared with those obtained with the same chromatin fraction upon depletion of linker histones and with the purified constitutive DNA counterpart. EXPERIMENTAL PROC...

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DNA sequence‐specific adducts of adriamycin and mitomycin C

Cited by 44 publications

References 25 publications

Characterization of covalent Adriamycin-DNA adducts

Characterization of covalent Adriamycin-DNA adducts

Mitomycinoid Alkaloids: Mechanism of Action, Biosynthesis, Total Syntheses, and Synthetic Approaches

Daunomycin-induced Unfolding and Aggregation of Chromatin

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