Abstract
Reactive compounds form adducts at several sites in DNA. One of these sites, the phosphate groups, forms phosphotriesters (PTE) which are both chemically stable and little repaired. A measurement of PTE in DNA could therefore be advantageous for the determination of doses in vivo of mutagens/cancer initiators. In this paper, the possibilities of utilizing the weakly alkylating properties of PTE for the transfer of adducts to strong nucleophiles have been investigated. Model compounds, thymidine 3′-[thymidine 5′-(methyl phosphate)], TpMeT, and thymidine 3′-[thymidine 5′-(2-hydroxyethyl phosphate)], TpHOEtT, were incubated with thiosulfate, a relatively strong nucleophile and the formation of dealkylated model PTE, thymidine 3′-(thymidine 5′-phoshate), TpT, was followed by HPLC. Transalkylation to thiosulfate or aniline of methyl PTE in DNA alkylated by [ 3H] N-methyl- N-nitrosourea was demonstrated. The methyl groups transferred, forming methyl thiosulfate and N-methylaniline, respectively, were determined by HPLC. These experiments demonstrate that it is possible to transfer alkyls from DNA phosphate to nucleophiles. Kinetic aspects of the transalkylation and requirement on nucleophiles for a practically useful method for determination of DNA adducts are discussed. Constants of reaction rates are presented.
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