Identification of paraldol-deoxyguanosine adducts in DNA reacted with crotonaldehyde

Crotonaldehyde (1) is a mutagen and carcinogen, but its reactions with DNA have been only partially characterized. In a previous study, we found that substantial amounts of 2-(2-hydroxypropyl)-4-hydroxy-6-methyl-1,3-dioxane ( paraldol, 7), the dimer of 3-hydroxybutanal (8), were released upon enzymat ic or neutral thermal hydrolysis of DNA that had been allowed to react with crotonaldehyde. We have now characterized two paraldol-deoxyguanosine addu cts in this DNA: N-2-[2-(2-hydroxypropyl)-6-methyl-1,3-dioxan-4-yl]deoxygua nosine (N-2-paraldol-dG, 13) and N-2-[2-(2-hydroxypropyl)-6-methyl-1,3-diox an-4-yl]deoxyguanylyl-(5'-3')-thymidine [N-2-paraldol-dG-(5'-3')-thymidine, 14]. Four diastereomers of N-2-paraldol-dG (13) were observed. Their overa ll structures were determined by H-1 NMR, by MS, and by reaction of paraldo l with deoxyguanosine and DNA. H-1 NMR data showed that two diastereomers h ad all equatorial substituents in the dioxane ring, while two others had an axial B-methyl group. Preparation of paraldol with the (R)- or (S)-configu ration at the 6-position of the dioxane ring and the carbinol carbon of the 2-(2-hydroxypropyl) group allowed partial assignment of the absolute confi gurations of N-2-paraldol-dG (13). Four diastereomers of N-2-paraldol-dG-(5 '-3')thymidine (14) were observed. Their overall structure was determined b y H-1 NMR, MS, and hydrolysis with snake venom or spleen phosphodiesterase. Reactions of nucleosides and nucleotides with paraldol demonstrated that a dducts were formed only from deoxyguanosine and its monophosphates. Experim ents with DNA that had been reacted with crotonaldehyde indicated that N-2- paraldol-dG-containing adducts in DNA are relatively resistant to enzymatic hydrolysis. The results of this study demonstrate that the reaction of cro tonaldehyde with DNA is more complex than previously recognized and that st able N-2-paraldol-dG adducts are among those that should be considered in a ssessing mechanisms of crotonaldehyde mutagenicity and carcinogenicity.