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  • Title: Inactivation of bacteriophage phi X174 by mitomycin C in the presence of sodium hydrosulfite and cupric ions.
    Author: Ueda K, Morita J, Yamashita K, Komano T.
    Journal: Chem Biol Interact; 1980 Feb; 29(2):145-58. PubMed ID: 6444372.
    Abstract:
    Bacteriophage phi X174 was inactivated by mitomycin C reduced with sodium hydrosulfite in the presence of cupric ions (Cu2+). 99% of the phage particles lost their plaque-forming abilities when incubated with 1.5 . 10(-4) M mitomycin C, 5.7 . 10(-4) M sodium hydrosulfite and 1.0 . 10(-4) M CuCl2 for 120 min at 37 degrees C in 0.05 M Tris--HCl buffer (pH 8.1). Sodium borohydride and thiol-reducing agents such as L-cysteine, 2-mercaptoethanol or dithiothreitol could not serve as a substitute for sodium hydrosulfite and other transition metal ions such as Fe2+, Fe3+, Mn2+, Co2+ and Zn2+ were of no effect. Inactivated phage sedimented at 114S just as intact phage, but phage DNA was degraded. Strand-scission was observed when phi X174 single-stranded DNA was directly reacted with mitomycin C reduced with sodium hydrosulfite in the presence of CuCl2. Phage inactivation was inhibited bycatalase, EDTA and several scavengers such as cysteamine, 2-aminoethylisothiuronium bromide HBr (AET), 4,5-dihydroxy-1,3-benzene-disulfonic acid (Tiron), or 1,4-diazabicyclo[2,2,2]octane (DABCO). These results suggest that free oxygen radicals and mitomycin C semiquinone radical generated during autoxidation of reduced mitomycin C in the presence of cupric ions cause the degradation of phy X174 DNA.
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