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Nucleic Acids Research, 1995, Vol. 23, No. 10 1664-1670
© 1995

CHEMISTRY

Novel reagents for chemical cleavage at abasic sites and UV photoproducts in DNA

Peter J. McHugh and John Knowland*

Department of Biochemistry, University of Oxford South Parks Road, Oxford 0X1 3QU, UK

* To whom correspondence should be addressed

Received February 27, 1995. Revised March 31, 1995. Accepted March 31, 1995.

Hot piperidine is often used to cleave abasic and UV-irradiated DNA at the sites of damage. It can inflict non-specific damage on DNA, probably because it Is a strong base and creates significant concentrations of hydroxyl ions which can attack purines and pyrimi-dines. We show that several other amines can cleave abasic DNA at or near neutral pH without non-specific damage. One diamine, N,N-dimethylethylenediamine, efficiently cleaves abasic DNA at pH 7.4 by either ß- or ß,{delta}-elimlnation, depending on temperature. Using end-labelled oligonucieotides we show that cleavage depends mainly on elimination reactions, but that 4',5'-cycllzation is also significant. This reagent also cleaves at photoproducts induced by (JVC and UVB, producing the same overall pattern as piperidine, but with no non-specific damage. It should prove valuable in locating low levels of photoproducts in DNA, such as those induced by natural sunlight.


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