Single Strand Scission Of DNA Research Articles

DFT investigations of phosphotriesters hydrolysis in aqueous solution: a model for DNA single strand scission induced by N-nitrosoureas

DNA phosphotriester adducts are common alkylation products of DNA phosphodiester moiety induced by N-nitrosoureas. The 2-hydroxyethyl phosphotriester was reported to hydrolyze more rapidly than other alkyl phosphotriesters both in neutral and in alkaline conditions, which can cause DNA single strand scission. In this work, DFT calculations have been employed to map out the four lowest activation free-energy profiles for neutral and alkaline hydrolysis of triethyl phosphate (TEP) and diethyl 2-hydroxyethyl phosphate (DEHEP). All the hydrolysis pathways were illuminated to be stepwise involving an acyclic or cyclic phosphorane intermediate for TEP or DEHEP, respectively. The rate-limiting step for all the hydrolysis reactions was found to be the formation of phosphorane intermediate, with the exception of DEHEP hydrolysis in alkaline conditions that the decomposition process turned out to be the rate-limiting step, owing to the extraordinary low formation barrier of cyclic phosphorane intermediate catalyzed by hydroxide. The rate-limiting barriers obtained for the four reactions are all consistent with the available experimental information concerning the corresponding hydrolysis reactions of phosphotriesters. Our calculations performed on the phosphate triesters hydrolysis predict that the lower formation barriers of cyclic phosphorane intermediates compared to its acyclic counter-part should be the dominant factor governing the hydrolysis rate enhancement of DEHEP relative to TEP both in neutral and in alkaline conditions.

Chartreusin, elsamicin A and related anti-cancer antibiotics.

Chartreusin and elsamicin A are structurally related antibiotics that bind to GC-rich tracts in DNA, with a clear preference for B-DNA over Z-DNA. They inhibit RNA synthesis and cause single-strand scission of DNA via the formation of free radicals. Elsamicin A can also be regarded as the most potent inhibitor of topoisomerase II reported so far. It can inhibit the formation of several DNA-protein complexes. Elsamicin A binding to the P1 and P2 promoter regions of the c-myc oncogene inhibits the binding of the Sp1 transcription factor, thus inhibiting transcription. Despite the pharmacological interest in chartreusin, elsamicin A and their derivatives, there is no experimental data on the structure of their complexes with DNA. This shortcoming has been partially solved by a theoretical approach, which provided some details about the DNA-elsamicin A interaction, and the thermodynamic characterization of the binding of chartreusin and elsamicin A to DNA. Elsamicin A but not chartreusin is being developed clinically as an anti-cancer agent. IST-622 (6-O-(3-ethoxypropylonyl)-3',4'-O-exo-benzylidene-chartreusin), a novel semi-synthetic derivative of chartreusin, which has shown a promising anti-cancer activity in a phase II study, appears to be a pro-drug with a more suitable pharmacokinetic profile than chartreusin.

Metabolic activation of o-phenylphenol to a major cytotoxic metabolite, phenylhydroquinone: role of human CYP1A2 and rat CYP2C11/CYP2E1

1. The in vitro metabolic activation of o-phenylphenol has been evaluated as yielding a toxic metabolite, 2,5-dihydroxybiphenyl (phenylhydroquinone), by p-hydroxylation in liver microsomes of rat and human. The involvement of rat CYP2C11, CYP2E1 and human CYP1A2 in the p-hydroxylation of o-phenylphenol is suggested. 2. 2,3- and phenylhydroquinone, which induced DNA single-strand scission in the presence of 1 μm CuCl2, were the most cytotoxic chemicals examined to cultured mammalian cell lines among o-phenylphenol, m-phenylphenol, p-phenylphenol, 2,2′-, 4,4′-, 2,3- and phenylhydroquinone. 3. Rat and human liver microsomes catalysed the formation of phenylhydroquinone, but not 2,3-dihydroxybiphenyl, using o-phenylphenol as a substrate. A higher rate of metabolic activation of o-phenylphenol was observed with livers of themale than the female rats by 5.6- and 2.6-fold respectively. 4. Inhibitory antibodies against the male-specific CYP2C11 inhibited hepatic o-phenylphenol p-hydroxylation in the male F344 and Sprague-Dawley rat by > 70%. Liver microsomes from the isoniazid-treated rats produced 1.8- and 3-fold induction of o-phenylphenol p-hydroxylation and chlorzoxazone 6-hydroxylation (a CYP2E1-dependent activity) respectively. 5. Human CYP1A2, expressed by baculovirus-mediated cDNA expression systems, exhibited a remarkably higher capacity for o-phenylphenol p-hydroxylation at concentrationsof 5 (> 5-fold), 50 (> 2-fold) and 500 μm (> 2-fold) than CYP2A, CYP2B, CYP2Cs, CYP2D6, CYP2E1 and CYP3A4 on the basis of pmol P450. 6. Among various CYP inhibitors tested here, 7,8-benzoflavone and furafylline, typical human CYP1A2 inhibitors, inhibited the microsomal p-hydroxylation of o-phenylphenol in human livers most potently by 70 and 50% respectively. 7. The results thus indicate the involvement of rat CYP2C11/CYP2E1 and human CYP1A2 in the hepatic p-hydroxylation of o-phenylphenol.

DNA single-strand scission in the pyloric mucosa of rat stomach induced by four glandular stomach carcinogens and three other chemicals

Induction of DNA single-strand scission by four glandular stomach carcinogens and three other chemicals was studied in the pyloric mucosa of rat stomach after gastric intubation. DNA single-strand scission, as was measured by the alkaline elution method, was induced by four glandular stomach carcinogens; N-nitroso- N-methylurethane at doses of 1 and 9 mg/kg body wt, 4-nitroquinoline 1-oxide at 20 and 30 mg/kg body wt, N-ethyl- N′-nitro- N-nitrosoguanidine at 30 and 100 mg/kg body wt and N-propyl- N′-nitro- N-nitrosoguanidine at 30 and 100 mg/kg body wt. DNA single-strand scission was also induced dose-dependently by a direct acting mutagen, 1-nitrosoindole-3-acetonitrile at doses of 100, 500 and 800 mg/kg body wt. Omeprazole, a proton pump inhibitor, was equivocal in its effect in this assay at 30–500 mg/kg body wt: induction was statistically significant by Cochran-Armitage binomial trend test. Loxitidine, an H 2-receptor antagonist, did not induce DNA single-strand breaks in the pyloric mucosa at a dose of 400 mg/kg body wt. The present results together with previous information suggest that SNA single-strand scission is a good marker for tumor-initiating activity in rat stomach mucosa.

Evidence for involvement of multiple iron species in DNA single-strand scission by H 2O 2 in HL-60 cells

Some of the properties of cellular iron species which react with H 2O 2 to cause DNA single-strand breaks in HL-60 cells were characterized in control cells and in cells made deficient of iron using 4,7-phenylsulfonyl-1,10-phenanthroline (bathophenanthroline disulfonic acid or BPS) and ascorbate. Single-strand breaks were measured using alkaline elution of DNA of cells treated at 4° to minimize repair during treatment. Strand breakage in the presence of 10% serum was only 40% of that in the absence of serum. This effect was traced to reaction of H 2O 2 with metals, most likely iron, in serum. Dimethyl sulfoxide (Me 2SO) inhibited a maximum of 65% of breaks in control cells. The diffusion distance from the site of generation of hydroxyl radicals to the site of reaction with DNA for the Me 2SO-inhibitable fraction was 6.9 nm. There was no significant alteration in the fraction of Me 2SO-inhibitable strand breaks or in diffusion distance in iron-deficient cells, though total strand breaks decreased by 70%. When the effect of extracellular iron in serum was taken into account, 60 μM orthophenanthroline (OP) inhibited a maximum of 85% of strand breaks. In cells pretreated with 60μM OP, the Me 2SO-inhibitable fraction of the remaining strand breaks decreased to 32%, while the diffusion distance decreased to 4.1 nm. These data indicate the existence of a number of different iron species, as characterized by overlapping but not coincidental inhibition by OP and Me 2SO, and by differing diffusion distances.

DNA damaging and cell proliferative activity of 1-methyl-1-nitrosourea in rat glandular stomach mucosa

The DNA damaging and cell proliferative activity of 1-methyl-1-nitrosourea (MNU), a glandular stomach carcinogen, was studied in the pyloric mucosa of male F344 rats after administration by gastric tube. DNA damage was measured with unscheduled DNA synthesis (UDS) and DNA single strand scission as markers, while cell proliferation was measured with replicative DNA synthesis (RDS) and ornithine decarboxylase (ODC) as markers. MNU at doses of 30 and 60 mg/kg body wt and 80 min after administration dose-dependently induced UDS (49 and 79 (0 dose, 19) dpm/μg DNA) measured by liquid scintillation counting in the presence of hydroxyurea (an inhibitor of RDS). RDS (DNA synthesis in the absence of hydroxyurea; 239 dpm/μg DNA at 0 dose) did not increase at that time. MNU at doses of 10 and 60 mg/kg body wt and 2 h after administration dose-dependently induced DNA single strand scission of 8.2 and 43.5 (0 dose, 1.4) elution rate constant (×10 −3/ml). MNU at doses of 30 and 60 mg/kg body wt and 24 h after administration dose-dependently induced an increase in RDS (1362 and 2393 (0 dose, 682) dpm/μg DNA). MNU at doses of 60, 90 and 120 mg/kg body wt and 24 h after administration dose-dependently induced an increase in ODC activity (22.0, 29.4 and 38.4 (0 dose, 6.3) p mol CO 2/30 min/mg protein). These results suggest that MNU has possible tumor initiating activity (UDS and DNA single strand scission) and tumor promoting activity (RDS and ODC) in rat stomach mucosa.

In vivo short-term assays for tumor initiation and promotion in the glandular stomach of Fischer rats

Here we summarize the data on 55 compounds tested in in vivo short-term assays for tumor-initiating and tumor-promoting activity in the glandular stomach of male Fischer (F344) rats. Most of the data has been previously published. Tumor-initiating activity was assayed by measuring the induction of unscheduled DNA synthesis (UDS) and DNA single strand scission; tumor-promoting activity was assayed by measuring the induction of ornithine decarboxylase (ODC) activity, increased replicative DNA synthesis (RDS), and of c- fos and c- myc oncogene expression. The compounds were orally administered. Twenty-nine compounds were tested for UDS. Eight were positive, including 5 glandular stomach carcinogens; 16 were negative, including 5 liver carcinogens; and 5 were equivocal. Twenty compounds were tested for DNA single strand scission. Twelve were positive, including 6 glandular stomach carcinogens; 7 negative, including 2 liver carcinogens; and 1 was equivocal. Thirty-two compounds were tested for RDS. Twenty-six were positive, including 8 glandular stomach carcinogens and 6 glandular stomach tumor-promoters; 4 were negative, including 3 liver carcinogens and a stomach irritant; and 2 were equivocal. Forty-five compounds were tested for ODC. Thirty-seven were positive, including 8 glandular stomach carcinogens and 6 glandular stomach tumor promoters; 7 were negative, including 3 liver carcinogens; and one was equivocal. All glandular stomach carcinogens and tumor-promoters examined were positive in both RDS and ODC. Two compounds were tested for c- fos and c- myc expression; one was a glandular stomach carcinogen and one was a glandular stomach tumor promoter, and both were positive. In addition, 2 compounds inhibited the increase in RDS induced by the tumor promoter NaCl, suggesting anti-tumor-promoter activity. Thus these assays are useful for assessing potential tumor-initiating and tumor-promoting activity in the rat glandular stomach.

Ability of N-methyl- N′-nitro- N-nitrosoguanidine, 4-nitroquinoline 1-oxide, dimethylnitrosome, and NaCl to induce inscheduled DNA synthesis, stimulate replicative DNA synthesis, and produce DNA single-strand breaks in pyloric mucosa of rat stomach

Male F344 rats were given test chemicals orally, and samples of their pyloric mucosa were incubated in vitro. Induction of unscheduled DNA synthesis (UDS) and stimulation of replicative DNA synthesis in the pyloric mucosa were then examined by addition of [ 3H]thymidine and simultaneous determinations of DNA synthesis in the presence of absence of hydroxyurea, an inhibitor of replicative DNA synthesis. DNA damage was also examined by the alkaline elution method with DNA single-strand scission as a marker. The results showed four types of abilities of the chemicals to effect UDS and replicative DNA synthesis in the pyloric of rat stomach 1–2 h after their administration: (1) induction of UDS and stimulation of replicative DNA synthesis by N-methyl- N′-nitro- N-nitrosoguanidine (MNNG), a glandular stomach carcinogen, (2) induction of only UDS by 4-nitroquinoline 1-oxide (4NQO), a glandular stomach carcinogen, (3) stimulation of only replicative DNA synthesis by NaCl, a glandular stomach tumor promoter, and (4) neither induction of UDS nor stimulation of replicative DNA synthesis by dimethylnitrosamine (DMN), a liver carcinogen. DNA single-strand scission was induced by MNNG and 4NQO, being maximal 2 h after their administration, but was not induced by NaCl or DMN. Thus it correlated well with the induction of UDS. The present results indicate four types of inductive abilities of chemicals on UDS and replicative DNA synthesis in rat stomach pyloric mucosa and show that this method can detect differences in the action mechanisms and organ specificities of glandular stomach carcinogens.

Possible tumor-initiating and -promoting activity of p-methylcatechol and methylhydroquinone in the pyloric mucosa of rat stomach.

The possible tumor‐promoting and tumor‐initiating activities of p‐methylcatechol and methylhydroquinone in the pyloric mucosa of male F344 rats were studied. Ornithine decarboxylase (ODC) and replicative DNA synthesis (RDS) were used as markers of tumor promotion and DNA single strand scission and unscheduled DNA synthesis (UDS) as markers of tumor initiation. The compounds were administered by gastric intubation and results were compared with those after administration of catechol. p‐Methylcatechol at doses of 60 to 180 mg/kg body weight dose‐dependently induced up to 20‐fold increase in ODC activity and 9‐fold increase in RDS with maxima 24 h after its administration, but it did not induce significant DNA single strand scission or UDS. Methylhydroquinone at a dose of 200 mg/kg body weight induced up to 6‐fold increase in ODC activity 24 h, and 5‐fold increase in RDS 16 h after its administration, but the induction was not dose‐dependent. At a dose of 200 mg/kg body weight it induced DNA single strand scission, but not UDS. These results and previous findings show that the possible tumor‐promoting activities of catechol are several times higher than those of p‐methylcatechol and 10 times higher than those of methylhydroquinone.

Determination of DNA single strand scission in stomach pyloric mucosa of rats treated with omeprazole and loxitidine

Determination of DNA single strand scission in stomach pyloric mucosa of rats treated with omeprazole and loxitidine

Genotoxicity and Cell Proliferative Activity of 3-Chloro-4-(Dichloromethyl)-5-Hydroxy-2(5H)-Furanone [MX] in Rat Glandular Stomach

MX is a strong direct acting mutagen on Salmonella typhimurium TA100 and is present in chlorinated tap water which contains organic compounds. MX was administered orally to 7-week-old male F344 rats, and its geno-toxicity in the pyloric mucosa of stomach was examined by analysis of DNA single strand scissions by the alkaline elution method. The effect of MX on cell proliferation was examined by assays of the inductions of replicative DNA synthesis and ornithine decarboxylase. MX at closes of 20-48 mg/kg body weight induced DNA single strand scissions dose-dependently (p<0.02) in the pyloric mucosa of the stomach 2 h after its administration. Moreover at doses of 10-60 mg/kg body weight, it induced up to 21-fold increase in replicative DNA synthesis (p<0.01) 16 h after its administration. At doses of 10-60 mg/kg body weight, it induced up to 100-fold increase in ornithine decarboxylase activity with a maximum 16 h after its administration. These results suggest that MX is genotoxic and induces cell proliferation in the glandular stomach of rats.

Genotoxicity and cell proliferative activity of a nitrosated Oroxylum indicum Vent fraction in the pyloric mucosa of rat stomach

In vivo genotoxic activity and cell proliferative activity were examined in the stomach mucosa of male F344 rats by in vivo short-term methods after oral administration of a nitrosated Oroxylum indicum Vent (OiV) fraction, which had been found to be mutagenic without S9 mix to Salmonella typhimurium TA98 and TA100. Administration of the nitrosated OiV fraction at doses of 1 and 2 g/kg body weight induced dose-dependent DNA single-strand scission ( p < 0.02), determined by the alkaline elution method, in the stomach pyloric mucosa 2 h after its administration: a dose of 2 g/kg body weight induced an 18-fold increase in the DNA elution rate constant. Administration of the nitrosated OiV fraction at doses of 0.7–2.8 g/kg body weight also induced dose-dependent increases, up to 100-fold ( p < 0.05), in replicative DNA synthesis in the stomach pyloric mucosa 16 h after its administration. Moreover administration of the nitrosated OiV fraction at doses of 0.25–2.0 g/kg body weight induced dose-dependent increases, up to 100-fold, in ornithine decarboxylase activity in the stomach pyloric mucosa with a maximum 4 h after its administration. These results demonstrate that the nitrosated OiV fraction has genotoxic and cell proliferative activity in the pyloric mucosa of rat stomach in vivo.

Covalent modification and single-strand scission of DNA by a new antitumor antibiotic kapurimycin A3

Kapurimycin A3 is a new antitumor antibiotic isolated from a Streptomyces. It contains the anthrapyrone skeleton and a beta,gamma-unsaturated delta-keto carboxylic acid moiety in the structure. In vitro, kapurimycin causes single-strand cleavage of supercoiled pBR322 DNA. The diminished cytotoxicity and DNA cleaving activity for 13-decarboxykapurimycin A3 indicates that the beta, gamma-unsaturated delta-keto carboxylic acid moiety is important for the activity of kapurimycin. Kapurimycin A3 binds to calf thymus DNA at 4 degrees C, and the thermal treatment of this adduct results in release of a guanine covalently attached to C-16 of kapurimycin via one of its nitrogen atoms. Thus, the epoxide is the alkylating functional group of kapurimycin, and this is consistent with the lack of DNA cleaving and cytotoxic activities for 14,16-deoxy-14,16-dihydroxykapurimycin. These findings have revealed that DNA strand scission by kapurimycin is due to the alkylation of guanine by ring opening of the epoxide group of kapurimycin, depurination of modified guanine, and presumably subsequent hydrolysis of the phosphate ester backbone at the resultant apurinic sites.

Inductions of Ornithine Decarboxylase and Replicative DNA Synthesis but not DNA Single Strand Scission or Unscheduled DNA Synthesis in the Pyloric Mucosa of Rat Stomach by Catechol

The possible tumor‐promoting and genotoxic activities of catechol were examined. Administration of catechol by gastric intubation at doses of 10 to 90 mg/kg body weight to male F344 rats induced up to 19‐fold increase in ornithine decarboxylase activity with a maximum after 8 h and up to 8‐fold increase in replicative DNA synthesis with a maximum after 24 h in the pyloric mucosa of the stomach. These results suggest that catechol has tumor‐promoting activity in the pyloric mucosa of rat stomach. However, its administration at doses of 37.5 to 90 mg/kg body weight did not induce DNA single strand scission in the pyloric mucosa as determined by the alkaline elution method after 2 and 6 h or unscheduled DNA synthesis examined after 2 and 12 h.

Site-directed mutagenesis of the T4 endonuclease V gene: role of tyrosine-129 and -131 in pyrimidine dimer-specific binding

T4 endonuclease V incises DNA at the sites of pyrimidine dimers through a two-step mechanism. These breakage reactions are preceded by the scanning of nontarget DNA and binding to pyrimidine dimers. In analogy to the synthetic tripeptides Lys-Trp-Lys and Lys-Tyr-Lys, which have been shown to be capable of producing single-strand scissions in DNA containing apurinic sites, endonuclease V has the amino acid sequence Trp-Tyr-Lys-Tyr-Tyr (128-132). Site-directed mutagenesis of the endonuclease V gene, denV, was performed at the Tyr-129 and at the Tyr-129 and Tyr-131 positions in order to convert the Tyr residues to nonaromatic amino acids to test their role in dimer-specific binding. The UV survival of repair-deficient (uvrA recA) Escherichia coli cells harboring the denV N-129 construction was dramatically reduced relative to wild-type denV+ cells. The survival of denV N-129,131 cells was indistinguishable from that of the parental strain lacking the denV gene. The mutant endonuclease V proteins were then characterized with regard to (1) dimer-specific nicking activity, (2) apurinic nicking activity, and (3) binding affinity to UV-irradiated DNA. Dimer-specific nicking activity and dimer-specific binding for both denV N-129 and N-129,131 were abolished, while apurinic-specific nicking was substantially retained in denV N-129,131 but was abolished in denV N-129. These results indicate that Tyr-129 and Tyr-131 positions of endonuclease V are at least important in pyrimidine dimer-specific binding and possibly nicking activity.

Involvement of DNA polymerases in the repair of DNA damage by benzo[ a]pyrene in cultured Chinese hamster cells

Mechanisms for induction of single-strand scissions in DNA by S9-activated benzo[ a]pyrene (B[a]P) and their repair in cultured Chinese hamster V79 cells were investigated with inhibitors of DNA-repair synthesis using alkaline sucrose gradient sedimentation analysis. The marked induction of single-strand scissions in DNA was observed following 3 h treatment of V79 cells with 5 μg/ml of B[a]P. These DNA lesions were repaired to the control level within 4 h after removal of B[a]P. The simultaneous addition of inhibitors of DNA-repair synthesis, 1-β- D-arabinofuranosylcytosine (araC) plus hydroxyurea with B[a]P did not increase the formation of DNA single-strand scissions. When these inhibitors were added after removal of B[a]P, however, they significantly blocked the rejoining of DNA-strand scissions. On the other hand, when aphidicolin, a specific inhibitor of DNA polymerase α, was used instead of araC, a partial inhibition of the rejoining was observed, and further addition of 2′,3′-dideoxythymidine, an inhibitor of DNA polymerase β, augmented the inhibitory effect. These results indicate that B[a]P-induced single-strand scissions of DNA in V79 cells could be repaired mostly by excision repair which involved DNA polymerase α and a non-α polymerase, presumably polymerase β.

Histochemical detection of DNA strand scissions in mammalian cells by [formula omitted] nick translation

A method to visualize in situ of single strand scissions of DNA in fixed mammalian cells has been developed. Using the nuclear nick translation with biotin-labeled dUTP followed by binding to avidin-biotin-peroxydase complex, the nuclei of HeLa cells which had been treated with a DNA-damaging antibiotic bleomycin were specifically stained, implicating that the histochemical detection of single strand scissions (nicks) of DNA in fixed cells was completed without destroying the morphology, and without using autoradiography.

REPAIR OF 8‐METHOXYPSORALEN INDUCED DNA INTERSTRAND CROSS‐LINKS IN Tetrahymena thermophila

Abstract— The protozoan Tetrahymena thermophila was treated with 8‐methoxypsoralen in combination with long wavelength ultraviolet irradiation and the DNA‐repair response was studied. Following the treatment a lag‐period in cell proliferation was observed, the duration of which was proportional to the amount of psoralen used, and both swelling and deformation of the cells were observed. The treatment with 3 μg 8‐methoxypsoralen/mℓ and a light dose of 8 kJ m‐2 resulted in a 10‐fold decrease in DNA and RNA synthesis activity, while the protein synthesis was only moderately affected. Using the same conditions the lag‐period was 30 h, and during this time the psoralen induced DNA interstrand cross‐links were removed. Alkaline elution experiments showed that the repair process involves DNA single strand scissions, whereas no double strand DNA scissions were detected.

Participation of active oxygen species in the induction of DNA single-strand scissions by cadmium chloride in cultured Chinese hamster cells

A mechanism for the induction of DNA single-strand scissions in cultured Chinese hamster cells by cadmium chloride (CdCl 2) was investigated by use of the technique of alkaline elution. Inducibility of DNA single-strand scissions by cadmium was examined under an aerobic or anaerobic culture condition. About 62% of the total cellular DNA was eluted throughout the filter within 10 h of elution time by treatment with 4 × 10 −5 M CdCl 2 for 2 h in our usual aerobic medium. In contrast, no difference in elution profiles of DNA was observed between untreated control cells and the cells treated with CdCl 2 in the anaerobic medium which was prepared by N 2 gas bubbling of aerobic medium for 60 min. Furthermore, elution of DNA from cells treated with cadmium decreased markedly in the presence of superoxide dismutase (SOD) when compared with that in the absence of SOD. Inhibition of the cell growth by cadmium was significantly protected by the presence of SOD in the medium although the cell growth was not restored to the control level. These results indicate that active oxygen species participate in Cd-induced DNA single-strand scissions and also in the growth inhibition of the cells by the metal.

Induction of 6-thioguanine-resistant mutants and single-strand scission of DNA by cadmium chloride in cultured Chinese hamster cells

Inducibility of 6-thioguanine-resistant (6TG r) mutants and single-strand scission of DNA by cadmium chloride (CdCl 2) was investigated in cultured Chinese hamster V79 cells. Frequency of 6TG r mutants increased concentration dependently by 24-h treatment with CdCl 2 up to 3 × 10 −6 M but decreased beyond 3 × 10 −6 M. Mutagenic potency of cadmium in the absence of S9 was about half that of benzo[ a]pyrene in the presence of S9 at equitoxic concentrations. Treatment of the cultured cells with cadmium after benzo[ a]pyrene treatment was not synergistic but additive to the mutagenicity of benzo[ a]pyrene. Single-strand scission of DNA by alkaline elution techniques was observed in the cells treated with CdCl 2 for 2 h in a concentration-dependent manner. The single-strand scission by cadmium was detected only in combination with proteinase K digestion of the cell lysates, indicating formation of DNA-protein cross-linking by the metal. These biological and biochemical findings indicate that cadmium is mutagenic in mammalian cells, and its mutagenic effect seems to be accompanied by single-strand scission of DNA.