Interaction of divalent and trivalent cations with antibiotic U-20,661, a DNA binding agent.
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Biomedical subjects
Publications and source records attributed to F Reusser.
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Antibiotic U-24,544 is a new agent isolated from the culture broth of a streptomycete strain. The antibiotic inhibits a variety of gram-positive and gram-negative bacteria in vitro, but is ineffective in treatment of experimental bacterial infections in mice. It is fairly cytotoxic in mammalian cell cultures and remarkably nontoxic in mice.
Antibiotic U-20,661 was shown to inhibit predominantly deoxyribonucleic acid (DNA)-directed ribonucleic acid (RNA) synthesis by binding to the double-stranded DNA template. Specific binding to DNA was verified by difference spectroscopy, reversal of the RNA polymerase inhibitory effect by increasing concentrations of DNA template, and by moderately increasing the melting temperature of double-stranded DNA in the presence of the antibiotic. The RNA polymerase reaction primed with synthetic poly dAT was inhibited considerably, but not completely even with high concentrations of antibiotic. Thus, the agent might bind to adenine or thymidine or both bases in the double-stranded DNA helix.
The effects of the three antibiotics U-12,241, nogalamycin, and U-20,661 on (i) deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) synthesis in KB cell cultures and cell-free systems of bacterial and mammalian origin and on (ii) oxidative phosphorylation in rat liver mitochondria were compared. Nogalamycin and U-12,241 inhibited RNA synthesis more strongly than DNA synthesis in all test systems. Antibiotic U-20,661 inhibited DNA and RNA synthesis equally in whole mammalian cells and their corresponding cell-free systems. The RNA polymerase from Escherichia coli, however, was at least 100 times more sensitive to U-20,661 than was the DNA polymerase. U-12,241 caused significant uncoupling of oxidative phosphorylation in mitochondria.
Antibiotic U-24,544, a new antibacterial agent, was found to be an effective uncoupler of phosphorylation associated with the oxidation of glutamate and succinate in rat liver mitochondria. Respiration was inhibited during glutamate oxidation but not during succinate oxidation. In a medium deficient in inorganic phosphate, the agent showed slight stimulation of mitochondrial glutamate oxidation. Mitochondrial swelling induced by inorganic phosphate was suppressed. The antibiotic inhibited protein, nucleic acid, and cell wall synthesis in Mycobacterium avium cells nearly equally without a predominant inhibition of any one of these macromolecular biosynthetic processes. Nucleic acid and polypeptide synthesis remained unaffected, but respiration was inhibited in cell-free bacterial systems. It was thus concluded that the antibiotic interfered primarily with the cellular energy-generating processes.
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