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Nucleic Acids Research, 1995, Vol. 23, No. 13 2450-2456
© 1995

CHEMISTRY

Use of oligonucletides to define the site of interstand cross-links induced by Adriamycin

Suzanne M. Cutts and Don R. Phillips*

School of Biochemistry, La Trobe University Bundoora, Victoria 3083, Australia

*To whom corespondence should be addressed

Received April 1, 1995. Accepted May 24, 1995.

It has been known for several years that Adriamycin forms adducts and interstrand cross-links when reacted for long periods of time with bacterial and mammalian DNA in vitro, with the cross-link being restricted to 2 bp elements containing GpC quences.The self-complementary 20mer deoxyoligonucleotide TA4T4GCA4T4A has been used in this study as a model of the apparent G-G cross-linking site at GpC sequences. The rate of formation of cross-links, as well as the dependence on both Adriamycin and Fe(lll) concentration, were similar with this llgonucleotlde as compared with calf thymus DNA. The cross-linking was demonstrated on both denaturing and non-denaturing sequencing gels. The half-life of the G-G cross-link was 40 h, consistent with that implied with high molecular weight, heterogeneous sequence DNA. Exonuclease III digests of adducts formed with 20mer eoxyoligonucleotides containing single, central G-G, G-I and l-l otential cross-links revealed that a guanine residue is required at both ends of the cross-link. No cross-linking was observed with a similar oligonucleotide containing only a single central (GC) bp.


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