DNA cross-links are biologically important because they compromise the function of cellular DNA. The pathways by which cells repair cross-links are important but are poorly understood. The preparation of chemically well-defined cross-linked DNA substrates for use in repair studies represents a significant challenge. Current methods involve multi-step syntheses and arduous purification protocols. An ideal method for the preparation of cross-linked DNA substrates for use in repair studies should involve a small number of simple chemical steps that are easily carried out in a biology or biochemistry laboratory, generate a single chemically well-defined species in good yield, and require little or no purification before use. Here we report the first method that meets these criteria. This method involves post-synthetic installation of a single hydrazino-dC residue (dC*) into a 2'-deoxyoligonucleotide, via treatment of a dC-containing oligonucleotide with bisulfite and hydrazine. A second 2'-deoxyoligonucleotide containing an abasic (Ap) site was then prepared by treatment of a 2'-deoxyuridine-containing oligonucleotide with the commercially available enzyme uracil DNA glycosylase (UDG). We present evidence that the hybridization of dC* and Ap containing oligonucleotides lead to rapid and high-yielding generation of interstrand cross-links presumably involving hydrazone formation between the aldehyde group of the Ap site and the hydrazine moiety of dC*.                                 

Preparation of DNA interstrand cross-links involving hydrazone formation between an abasic site and a N<sup>4</sup>-aminocytidine residues in duplex DNA

248th National Meeting (2014)

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Preparation of DNA interstrand cross-links involving hydrazone formation between an abasic site and a N4-aminocytidine residues in duplex DNA

Preparation of DNA interstrand cross-links involving hydrazone formation between an abasic site and a N⁴-aminocytidine residues in duplex DNA