Nucleic acid-specificity of bleomycin

TY - JOUR

T1 - Nucleic acid-specificity of bleomycin

AU - Haidle, Charles W.

AU - Kuo, Macus Tien

AU - Weiss, Karolyn Kay

N1 - Funding Information: BLEOMYCIN causes a decrease in the melting temperature of DNA and, in addition, produces a scission of DNA strands both in vitro and in vivo, the former reaction requiring 2-mercaptoethanol or hydrogen peroxide.1-3 In vitro, high concentrations of the drug render about 80 per cent of the DNA trichloroacetic acid (TCA) soluble4 and also cause the release of free bases from DNA.’ In addition to the scission of native DNA strands, Nagai et al.3 showed that bleomycin also caused the fragmentation of the synthetic deoxyribopolymers, polydeoxycytidylate and polydeoxyguanidyl-ate. However, Suzuki et aL6 observed no fragmentation of ribosomal or transfer RNA in the presence of bleomycin. The oresent studv was initiated to determine the reason for the apparent specificity of bleomycin toward-DNA. _ The bleomycin was a gift from Bristol Laboratories, Syracuse, N.Y. (lot No. 701233), and from Niooon Kavaku Comoanv. Tokyo. Japan, (lot No. F7071BS). This bleomycin has an absorption maximum at 292 nm. i]ri&e-2-‘d (5OIO& /m-mole) was purchased from International Chemical and Nuclear Corp., Irvine, Calif. Polyuridylate-3H (78.1 mCi/m-mole phosphorus) and the copolymer, polydeoxyandenylate-thymidylate-3H (25.1 mCi/m-mole phosphorus) were purchased from Miles Laboratories, Inc., Elkhart, Ind. Uracil-2-14C (56.3 mCi/m-mole) and uridine-5-3H (27.8 Ci/m-mole) were purchased from New England Nuclear, Boston, Mass. Polyadenylate-3H (51.0 mCi/ m-mole phosphorus), thymidinemethyl-3H (11.9 Ci/m-mole) and thymine-2-l% (48.6 mCi/m-mole) were purchased from Schwarz BioResearch, Orangeburg, N.Y. Bovine pancreatic ribonuclease A (EC 2.7.7.16), 5 times crystallized, protease-free, was purchased from Sigma Chemical Company, St. Louis, MO. Yeast ribonucleic acid sodium salt was purchased from Mann Research Laboratories, Inc., New York, N.Y. All other chemicals were purchased from commercial sources. Labeled bacterial DNA was prepared from Bacillus subtifis 168 (thy trp C2) as previously described.“ Uridine-5-3H-labeled DNA, in which deoxyuridylate replaces thymidylate,6 was prepared from confluent lysis plates of bacteriophage PBS-l using B. subtilis 168 (trp C2) as the host bacterium. The plating procedure of Takahashi was used with minor modifications.7-9 Labeled RNA was prepared from B. subtilis (thy trp C2) according to the method of Okamoto et ~1.‘~T he bacterial cells were grown as previously described for the isolation of DNA, except that 0.2 &i/ml of uridine-2 I‘% was added to the culture medium. DNA concentration was assayed by the diphenylamine method of Burton.” RNA concentration was assayed by the orcinol method of Hurlbert et al.” * This research was supported in part by Grant CA 13246 from the United States Public Health Service and Grant G-441 from the Robert A. Welch Foundation.

PY - 1972/12/15

Y1 - 1972/12/15

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U2 - 10.1016/0006-2952(72)90096-2

DO - 10.1016/0006-2952(72)90096-2

M3 - Article

C2 - 4119299

AN - SCOPUS:0015511677

SN - 0006-2952

VL - 21

SP - 3308

EP - 3312

JO - Biochemical Pharmacology

JF - Biochemical Pharmacology

IS - 24

ER -