Selective Abstraction of 2H from C-5' of Thymidylate in an Oligodeoxynucleotide by the Radical Center at C-6 of the Diradical Species of Neocarzinostatin: Chemical Evidence for the Structure of the Activated Drug-DNA Complex

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1991 Apr 15

PMID 1826561

Citations 4

Authors

S M Meschwitz

I H Goldberg

Affiliations

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Abstract

Use has been made of the mechanism of DNA deoxyribose damage by the ene-diyne-containing chromophore of the antitumor antibiotic neocarzinostatin to provide chemical evidence for the structure of the activated drug-DNA complex. Radical centers at C-2 and C-6 of the diradical form of the glutathione-activated chromophore abstract hydrogen atoms from C-1' of the C residue and C-5' of the T residue in AGC.GCT to generate a bistranded lesion consisting of an abasic site at C and a strand break at T. This laboratory has proposed a molecular model for the drug-DNA interaction in which the naphthoate moiety of the chromophore intercalates between A.T and G.C, placing the diradical core in the minor groove, so that the radical centers at C-6 and C-2 are close to C-5' of T and C-1' of C, respectively. To determine which radical center abstracts one of the hydrogen atoms from C-5', the self-complementary oligodeoxynucleotide GCAGCGCTGC was synthesized with 2H at both 5' positions of the T residue and treated with glutathione-activated chromophore. Sequencing-gel electrophoresis showed that drug attack was limited to the T and C residues and that abstraction of 2H from C-5' exhibited a small isotope selection effect of 1.25. 1H NMR spectroscopic examination of the reacted chromophore, isolated by HPLC, indicated that 2H was selectively abstracted by C-6, providing experimental corroboration of the model and further elucidating the chemical mechanism. Since direct strand breakage at the T residue exceeds (44% more) abasic site formation at the C residue, other models of drug-DNA interaction leading to only single-strand breaks are also considered.

Citing Articles

Reactivity of the Major Product of C5'-Oxidative DNA Damage in Nucleosome Core Particles.

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Modulation of neocarzinostatin-mediated DNA double strand damage by activating thiol: deuterium isotope effects.

McAfee S, Ashley G Nucleic Acids Res. 1992; 20(4):805-9.

PMID: 1531872 PMC: 312021. DOI: 10.1093/nar/20.4.805.

Neocarzinostatin acts as a sensitive probe of DNA microheterogeneity: switching of chemistry from C-1' to C-4' by a G.T mismatch 5' to the site of DNA damage.

Kappen L, Goldberg I Proc Natl Acad Sci U S A. 1992; 89(15):6706-10.

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