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Nucleic Acids Research, 1985, Vol. 13, No. 2 595-603
© 1985

Articles

Kinetic evidence that echinomycin migrates between potential DNA binding sites

Keith R. Fox and Michael J. Waring

University of Cambridge Department of Pharmacology, Medical School Hills Road, Cambridge CB2 2QD, UK

Received October 26, 1984. Revised December 13, 1985. Accepted December 13, 1985.

The hypothesis that echinomycin locates its preferred nucleotide sequences in DNA by a process of "shuffling" between potential binding sites has been tested. Immediately after reacting with calf thymus DNA the antibiotic is relatively weakly bound inasmuch as the complex dissociates quite rapidly when detergent is added. If the complex is allowed to equilibrate for various periods of time after mixing, an increasing proportion of the bound antibiotic dissociates slowly on addition of detergent. The kinetics of appearance of the slowly-dissociating form, and its dependence upon ionic strength, are fully consistent with the shuffling model. In contrast the dissociation profiles from poly(dG–dC) and poly(dA–dT) are independent of mixing time.


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