Photofootprinting of drug-binding sites on DNA using diazo- and azido-9-aminoacridine derivatives |
| |
Authors: | C Jeppesen P E Nielsen |
| |
Institution: | Department of Biochemistry B, Panum Institute, University of Copenhagen, Denmark. |
| |
Abstract: | It is demonstrated that DNA photofootprinting analysis of the intercalating depsipeptide echinomycin, and the minor groove-binders distamicyn, 4',6-diamidino-2-phenylindole (DAPI) and Hoechst 33258 can be performed using 9-6-(2-diazocyclopentadienylcarbonyloxy)hexylamino]acridine (DHA) Nielsen et al. (1988) Nucleic Acids Res. 16, 3877-3888] or 2-methoxy-6-azido-9-aminoacridine (MAA) Jeppesen et al. (1988) Nucleic Acids Res. 16, 5755-5770]. Both the extent of the drug-binding sites and their relative strength can be determined with either reagent. DNA has the advantage of giving virtually sequence-uniform DNA photocleavage. On the other hand, structural changes in the DNA are detected by MAA. Using the 232-base-pair EcoRI-PvuII pUC19 restriction fragment, it is found that cleavage protection by distamycin, DAPI and Hoechst 33258 all require an (A.T)4 sequence, whereas protection by echinomycin was confined to a G + C-rich 8-base-pair region. |
| |
Keywords: | |
|
|