Detection of minor adducts in cisplatin-modified DNA by transcription footprinting

Two DNA restriction fragments containing either a d(GC)5 or a d(TTGCTTGATTAGTTGTGTT) insert were subjected to reaction with cis-diamminedichloroplatinum(II) and were then used as templates for RNA synthesis by T7 RNA polymerase. Within the d(GC)5 insert, interstrand cross-links are preferentially formed. Within the second insert, the reactivity order of the potential binding sites is d(ApG) > d(GpC/GpC) = d(GpA) > d(GpTpG). In the presence of cyanide ions, the adducts are much less stable at the d(GpA) sites than at the d(GpCpG) sites, in double-stranded DNA.