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A High-Throughput Comet Assay Approach for Assessing Cellular DNA Damage
Published on: May 10, 2022
The effect of chromatin structure on cisplatin damage in intact human cells
N P Davies1, L C Hardman, V Murray
1School of Biochemistry and Molecular Genetics, University of New South Wales, Sydney, NSW 2052, Australia.
Abstract:
The influence of chromatin structure on cisplatin DNA damage was investigated in intact human cells. The epsilon-globin gene promoter was utilised as the target DNA sequence and the terminal transferase-dependent PCR technique was employed to examine adduct formation at base pair resolution. It was found that cisplatin preferentially damaged at runs of consecutive guanine bases in intact cells. By comparing the relative intensity of adduct formation in intact cells and in purified genomic DNA, it was possible to assess the influence of chromatin proteins on the extent of cisplatin DNA damage. Enhanced damage in intact cells was found at the CACC site where a member of the Sp1 family of proteins is thought to bind. It is postulated that protein binding at this site bends the DNA double-helix so that enhanced cisplatin binding occurs. The altered DNA binding of cisplatin in the presence of chromatin proteins could be important in the properties of cisplatin as an anti-tumour drug.
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