Characterization of Bleomycin-Resistant DNA

After reaction of DNA with high concentrations of bleomycin, approximately 80% of the DNA becomes trichloroacetic acid (TCA) soluble. The remaining 20% of the DNA remains TCA insoluble. Upon further treatment of this TCA-insoluble material with high concentrations of the drug, no further drug action...

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Bibliographic Details
Published in:Biophysical journal 1973-12, Vol.13 (12), p.1296-1306
Main Authors: Kuo, M.Tien, Haidle, Charles W., Inners, L.Daniel
Format: Article
Language:English
Subjects:
Animals
Bacillus subtilis - analysis
Binding Sites
Bleomycin
Cell Line
Centrifugation, Density Gradient
Coliphages - analysis
Cricetinae
Deoxyribonucleases
DNA - analysis
DNA, Bacterial - analysis
DNA, Single-Stranded - analysis
DNA, Viral - analysis
Drug Resistance
Hot Temperature
Lung - analysis
Molecular Weight
Nucleic Acid Denaturation
Pancreas - enzymology
Phosphoric Diester Hydrolases
Phosphorus Radioisotopes
Solubility
Thymidine
Trichloroacetic Acid
Tritium
Ultracentrifugation
Venoms
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Summary:After reaction of DNA with high concentrations of bleomycin, approximately 80% of the DNA becomes trichloroacetic acid (TCA) soluble. The remaining 20% of the DNA remains TCA insoluble. Upon further treatment of this TCA-insoluble material with high concentrations of the drug, no further drug action can be detected. Drug action is defined as fragmentation of DNA to smaller molecular size, release of free bases, and TCA solubilization. This material which is not attacked by bleomycin has been termed bleomycin-resistant DNA. This bleomycin-resistant DNA does not compete with native DNA in the bleomycin reaction indicating that there is no binding or inactivation of the drug by the resistant DNA. The resistant DNA shows very little hyperchromicity when heated through the melting temperature for the corresponding native DNA, indicating a single-stranded structure. Results of sedimentation and equilibrium analyses yield a molecular weight of about 4,000 daltons. This value is the same regardless of the source of the native DNA. Finally, the bleomycin-resistant DNA exhibits a base composition similar to that of the native DNA from which it was derived.
ISSN:0006-3495
1542-0086
DOI:10.1016/S0006-3495(73)86063-1