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Effect of apurinic/apyrimidinic endonucleases and polyamines on DNA treated with bleomycin and neocarzinostatin: specific formation and cleavage of closely opposed lesions in complementary strands

Journal Article · · Biochemistry; (United States)
DOI:https://doi.org/10.1021/bi00410a049· OSTI ID:7024779
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Bleomycin and neocarzinostatin induce modified apurinic/apyrimidinic (AP) sites by oxidation of the sugar moiety in DNA. In order to quantitatively assess the susceptibility of these lesions to repair endonucleases, drug-treated /sup 3/H-labeled colE1 DNA was mixed with /sup 14/C-labeled heat-depurinated DNA, and endonuclease-susceptible sites in the mixture were titrated with various AP endonucleases or with polyamines. Single- and double-strand breaks were quantitated by determining the fractions of supercoiled, nicked circular, and linear molecules. Exonuclease III and endonucleases III and IV of Escherichia coli, indicating cleavage of drug-induced AP sites. Bleomycin-induced AP sites were much more sensitive to cleavage by putrescine than heat-induced sites. Treatment with putrescine or very high concentrations of endonuclease III also increased the number of double-strand breaks in bleomycin-treated DNA, suggesting a minor class of lesion consisting of an AP site accompanied by a closely opposed break in the complementary strand. These complex lesions were resistant to cleavage by endonuclease IV. These results suggest that virtually all neocarzinostatin-induced AP sites are accompanied by a closely opposed strand break. Several characteristics of the putative AP site/strand break lesions induced by neocarzinostatin suggest that they may correspond to certain AP-like lesions which were previously detected on DNA sequencing gels as endonuclease IV susceptible sites and which have been strongly implicated in neocarzinostatin-induced mutagenesis.

Research Organization:
Virginia Commonwealth Univ., Richmond (USA)
OSTI ID:
7024779
Journal Information:
Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 27:10; ISSN BICHA
Country of Publication:
United States
Language:
English

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Related Subjects

550201 -- Biochemistry-- Tracer Techniques
560300* -- Chemicals Metabolism & Toxicology
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AMINES
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
ANTIMITOTIC DRUGS
ANTINEOPLASTIC DRUGS
BACTERIA
BIOCHEMISTRY
BIOLOGICAL EFFECTS
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
BLEOMYCIN
CARBON 14 COMPOUNDS
CHEMISTRY
DNA
DNA REPAIR
DNA REPLICATION
DNA-ASE
DRUGS
ENDONUCLEASES
ENZYMES
ESCHERICHIA COLI
ESTERASES
HYDROLASES
LABELLED COMPOUNDS
MICROORGANISMS
NEOCARCINOSTATIN
NUCLEIC ACID REPLICATION
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PHOSPHODIESTERASES
RADIOMIMETIC DRUGS
RECOVERY
REPAIR
STRAND BREAKS
TRITIUM COMPOUNDS