Light-induced strand scission of DNA by cobalt-bleomycins is more likely to occur at certain base sequences than others. By use of 32P-end-labeled DNA restriction fragments as the substrates for cleavage, products have been analyzed on high-resolution polyacrylamide gels and compared to those produced by iron-bleomycin. The results indicate that the sites of damage to DNA are similar in both cases: pyrimidine residues located at the 3′ side of a guanine are preferentially attacked. Consistent with the observed nicking specificity, interactions between cobalt-bleomycin and guanine residues in the trinucleotide sequence GGT are revealed in a dimethyl sulfate methylation experiment. The action spectrum for the light-induced DNA cleavage reaction correlates with the absorption spectrum of cobalt-bleomycin in the wavelength range between 330 and 450 nm. In contrast to iron-bleomycin, the extent of DNA degradation by light-activated cobalt-bleomycins appears to be indifferent to the concentration of dissolved oxygen in the reaction medium, and little or no base propenal is produced. Bases (e.g., thymine) are released by both agents. Fluorescence quenching experiments show that apparent binding constants of cobalt-bleomycin complexes with DNA are in the 107 M-1 range in 25 mM tris(hydroxymethyl)aminomethane, pH 8, 1 mM NaCl, and 1 mM ethylenediaminetetraacetic acid at 25 °C.
|Original language||English (US)|
|Number of pages||7|
|State||Published - 1984|
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