Detection of Platinum Lesions at the Nucleotide Level in Cells using Single Strand Ligation PCR
The mechanism of action of many anti-cancer drugs, including platinum based drugs, involves direct, covalent adduct formation on nucleotides in DNA. Cisplatin forms mainly intrastrand cross-links between two adjacent guanines in runs of two or more consecutive guanines and between AG pairs.1,2 Recently, the sequence preferences of lesion formation have been studied in isolated DNA3 by exploiting the finding that such lesions can block the progression of DNA taq polymerases. This and other assays using plasmid DNA have revealed that even relatively simple DNA damaging agents such as cisplatin show a degree of sequence preference in adduct formation.4,5 Studying drug interactions in artificial systems with oligonucleotides or with plasmid DNA provides useful information but cannot necessarily predict intracellular behaviour where DNA exists in a highly ordered structure, complexed with many proteins, and where other cellular components may affect reactivity.
KeywordsSingle Copy Gene Sequence Preference Intrastrand Crosslinks Consecutive Guanine Covalent Adduct Formation
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