Isolation and Characterization of an Interstrand Cross-Link Adduct Between Aziridinomitosenes and DNA

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Student Presentation

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Don Warner


Mitomycin C, a clinically used chemotherapeutic agent, forms interstrand covalent cross-links in the minor grove of DNA at CG•CG specific sites. The DNA cross-links are the source of mitomycin C’s cytotoxicity and antitumor activity, resulting from their obstruction of DNA replication. The reductive activation process for mitomycin C, which makes it biologically active, produces oxygen radicals in the body that potentially contribute to its toxic side effects. Aziridinomitosenes follow a similar mechanism of interaction, but have the potential to work under non-reductive conditions, thus potentially reducing the toxic side effects. To better understand the mechanism by which aziridinomitosenes cross-link DNA, we sought to isolate and characterize the bisadduct between UMMS, a specific aziridinomitosene, and the two guanine bases at CG•CG specific sites within duplex DNA. The method employed first required the incubation of UMMS with a synthetic self-complimentary oligonucliotides to form the DNA cross-links. The cross-linked material was then isolated using denaturing polyacrylamide gel electrophoresis. The purified cross-linked DNA was then enzymatically digested and the reaction mixture was analyzed with LC/MS. The isolation and characterization of the bisadduct between UMMS and DNA will aid in the understanding of both the cytotoxic and antitumor potential of aziridinomitosenes.

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