Modification of natural, double-helical DNA by antitumor cis- and trans-[Cl-2(Me2SO4)(4)Ru] in cell-free media
Modifications of natural DNA in cell-free media by the antitumor ruthenium compounds cis- and trans-[Cl-2(Me2SO4)(4)Ru] were studied by various biochemical and biophysical methods. These methods included: binding studies by means of flameless atomic absorption spectrophotometry, mapping of DNA adducts by means of transcription assay, use of ethidium bromide as a fluorescent probe of DNA adducts of metal complexes, an interstrand cross-linking assay employing gel electrophoresis under denaturing conditions, measurements of DNA unwinding by ger electrophoresis, differential pulse polarographic analysis of DNA conformation, and analysis of liquid crystalline dispersions of DNA by circular dichroism. The results indicated that both ruthenium compounds irreversibly coordinated to DNA; the rate of binding of the cis isomer was considerably lower than that of the trans isomer. The DNA-binding mode of trans-[Cl-2(Me2SO4)(4)Ru] included formation of bifunctional adducts such as intrastrand cross links between neighboring purine residues and a small amount (similar to1%) of interstrand cross-links. cis-[Cl-2(Me2SO4)(4)Ru] formed mainly monofunctional lesions on natural DNA. Both ruthenium isomers induced conformational alterations of non-denaturational character in DNA, the trans compound being more effective. In addition, DNA adducts of trans-[Cl-2(Me2SO4)(4)Ru] were capable of inhibiting RNA synthesis by DNA-dependent RNA polymerases, while the adducts of the cis isomer were not. Thus, several features of the DNA-binding mode of trans-[Cl-2(Me2SO4)(4)Ru] were similar to those of antitumor cis-diamminedichloroplatinum (II), which may be relevant to the biological effects of this antitumor ruthenium drug. On the other hand, the different DNA-binding mode of cis-[Cl-2(Me2SO4)(4)Ru] was consistent with its less pronounced biological effects. BIOCHEM PHARMACOL 60;12:1761-1771, 2000. (C) 2000 Elsevier Science Inc.