Nickel induces a signature mutation for oxygen free radical damage.

We have determined the specificity of mutations produced by nickel(II), a known human carcinogen, in a forward mutation assay and also used a sensitive reversion assay to show that Ni(II), like iron and copper, can produce tandem double CC-->TT mutations, a hallmark of damage to DNA by either UV irradiation or oxygen free radicals. A reduction in mutation frequencies by the addition of oxygen radical scavengers also supports the involvement of reactive oxygen species in DNA damage and mutagenesis by Ni(II). Mutagenesis by Ni(II) is enhanced by the addition of both hydrogen peroxide and a tripeptide glycyl-glycyl-L-histidine. The enhancement of mutagenesis of Ni(II) by the tripeptide indicates that these complexes could serve to localize Ni(II) in nuclei and mediate DNA damage and mutagenesis via the generation of short-lived oxygen free radicals. These data suggest that Ni(II) carcinogenesis may proceed via the generation of active oxygen species and furthermore provide a model for nickel carcinogenesis based on the binding of Ni(II) to nuclear proteins.

Effects of arsenic, selenium, and chromium on the fidelity of DNA synthesis.

The effect of three environmentally important metals, arsenic, selenium, and chromium, on the accuracy of DNA synthesis in vitro has been analyzed. The addition of arsenic to fidelity assays did not significantly alter accuracy. Selenium did not alter fidelity under normal conditions of magnesium activation, nor did it affect the mutagenicity of manganese. Chromium in the form of Cr(III) as well as Cr(VI) diminished the fidelity by which Escherichia coli DNA polymerase I copies polynucleotide templates. Nearest-neighbor analysis of the product synthesized in the presence of chromium indicates that the misincorporated in the presence of chromium indicates that the misincorporated bases are present as single-base substitutions. Chromium was also mutagenic using the recently developed phi chi 174 assay, which measures the fidelity of DNA synthesis with a natural DNA template.

Metal-induced infidelity of DNA synthesis.

A number of metals have been demonstrated to be mutagens in procaryotic and eucaryotic organisms as well as carcinogens in experimental animals. Epidemiologic studies have indicated that Ni, Cr, and As are involved in human carcinogenesis. We have hypothesized that the active molecular species is the cation and that metal induced mutations result from incorrect base-substitutions during DNA replication. This is supported by the observations that metal ions diminish the fidelity of DNA synthesis in vitro using a variety of DNA polymerases. There is a significant correlation between the metals that decrease fidelity and those that have been reported to be mutagenic and carcinogenic. Thus, metal carcinogens are no exception to the general postulate that carcinogens can be identified by their effects on DNA.

Metal-induced lethality and mutagenesis: possible role of apurinic intermediates.

The possible mechanisms by which various metals exert their mutagenic effects were investigated using both chemical and biochemical techniques. Ions of Cu, Ni and Cr enhanced the release of either adenine [Cu(II) and Ni(II)] or guanine [Cr(VI)] from DNA as measured in a chromatography assay, suggesting the possible importance of depurination in metal-induced mutagenesis. Transfection experiments with single-stranded bacteriophage phi X174 DNA indicated that micromolar levels of Cu(II), Cr(III), Cr(VI) and Pt(IV) are capable of causing extensive lethal damage to the phage DNA. In case of Cu(II) and Pt(IV) this damage proved mutagenic for phi X174 am3 after transfection of DNA into SOS-induced spheroplasts. For Cu(II) mutagenesis is likely due to the release of adenine residues from the phage DNA based on the abolishment of mutagenesis by alkali and the observed specificity of the phage revertants. The enhancement of the adenine depurination rate by Cu(II) was estimated to be as high as 10,000-fold.

Effect of some nickel compounds on red blood cell characteristics.

The effect of certain inorganic and coordinated nickel compounds on the resistance to different destructive substances, rheological properties, and functional activity of healthy human red blood cells (RBC), was investigated. It is shown that nickel compounds affect the erythrocyte membrane lipid bilayer, as well as membrane proteins to various extents, depending on the type of compounds used. In general, the acceleration of erythrocyte aging was observed to be more pronounced in young erythrocytes. The observed results suggest that nickel compounds decrease water permeability across erythrocyte membranes. Almost all the investigated nickel compounds decrease erythrocyte thermostability, deformability, and the rate of O2 release by erythrocytes.

[Comparative study of nucleosome particles in chromatin from normal and tumor cells. I. Structural parameters]. / Sravnitel'noe issledovanie nukleosomnykh chastits khromatina normal'nykh i opukholevykh kletok. I. Strukturnye parametry.

The composition and structure of nucleosomic fragments isolated from the ascitic hepatoma 22A cells, liver and from cells of C3HA mice in norm and after partial hepatectomy were investigated. Via electrophoresis in 1.5% agarose gel with the emplogment of reperic restrictive DNA fragments and with the help of mathematical processing, the value of the nucleosomic DNA repeat in ascitic hepatoma 22A was calculated to be 187 b.p., and in regenerating liver--196 b.p. The absence of the H1 degree subfraction in chromatin of ascitic hepatoma 22A cells was found. Lower electrophoretic mobility in 5% polyacrylamid gel of nucleosomic chromatin fragments of ascitic hepatoma 22A as compared with their counterparts from healthy mice liver was established. The method of circular dichroism allowed to reveal differences in the RNA and protein structural state in nucleosomes of normal and tumour cells. The structure of nucleosomes of regenerating mice liver of the C3HA strain did not differ from that of normal liver of the same mice.

[Concentration of DNA divalent ions in transformed tissues during exposure to various factors]. / Soderzhanie dvukhvalentnykh ionov DNK iz transformirovannykh tkanei pri razlichnykh vozdeistviiakh.

The effect of X-irradiation (1000 r) on the rat liver nuclear DNA Zn content has been studied. The results show a significant decrease of Zn content 1 hour after irradiation and some its normalization after 24 hours. The obtained data on Zn content decrease may be considered as one of the responses of nuclear DNA to irradiation.

[Ca2+, Mg2+, Zn2+ levels in histone fractions from normal and tumor cells]. / Soderzhanie Ca2+, Mg2+, Zn2+ vo fraktsiiakh gistonov normal'nykh i opukholevykh kletok.

Distribution of some bivalent cations (Ca2+, Mg2+, Zn2+) in histones isolated from healthy mice liver and ascitic hepatoma 22A cells has been investigated by atomic-absorption analysis. It has been shown that the content of these cations is higher in normal and diseased H3, H2B and H1 fractions and lower--in H2A; however, in the H4 fraction these metals are not detected. A significant increase of Ca2+, Mg2+ and Zn2+ levels has been established in ascitic H3, H2B and H1 fractions. An increase of bivalent cations (Ca2+, Mg2+, Zn2+) content in some histone fractions apparently is bound with the changes of histone--histone and histone--DNA interactions.

Mutation spectrum of copper-induced DNA damage.

The ability of metal ions to damage DNA and cause mutagenesis has been analyzed with reversion and forward mutation assays using single-stranded DNA templates. We previously reported that incubation of phi X174 am3 DNA with Fe2+ in vitro results in mutagenesis when the treated DNA is transfected into Escherichia coli spheroplasts (Loeb, L. A., James, E. A., Waltersdorph, A. M., and Klebanoff, S. J. (1988) Proc. Natl. Acad. Sci. U.S.A. 85, 3918-3922, 1988). We now extend these studies to other metal ions. Of the metal ions tested, copper ions were the most mutagenic; the frequency of mutants produced was equal to or greater than that produced by Fe2+. Mutagenesis by Cu+ was diminished by catalase, mannitol, and superoxide dismutase suggesting the involvement of H2O2, hydroxyl ions, and superoxide, respectively. However, the findings that Cu+ and Cu2+ are nearly equally mutagenic and that the mutagenic activities are not completely inhibited by oxygen free radical scavengers make it unlikely that the mechanism for mutagenesis is simply the production of hydroxyl free radicals. The spectra of mutations produced by either copper ion using the lacZ gene as a target are very similar and differ from those reported with other agents. The predominant mutagenic sequence changes are single-base substitutions, the most frequent being replacement of a template C by a T. This transition presumably results from mispairing of an altered C with deoxyadenosine. Copper-induced mutations are not randomly distributed. Instead, they are found predominantly in clusters suggesting direct interaction of copper ions with specific nucleotide sequences in DNA. Evidence is considered that the high frequency of C----T transitions may be a common manifestation of DNA damage by oxygen radicals.

[Possibility of migration of several elements in biological systems following x-irradiation]. / O vozmozhnosti migratsii nekotorykh élementov v biologicheskikh sistemakh pri rentgenovskom obluchenii

The effect of local X-irradiation on the content of trace elements of subcellular fractions (nuclear, mitochondrial) and the blood serum of rats with sarcoma M-1 was studied. It was shown that systematic local irradiation of sarcoma M-1 (single dose--250 R) during the tumour growth promoted zinc redistribution from the nuclei into the mitochondria. In addition, there were revealed statistically significant postirradiation changes in the blood serum Ca content of rats with sarcoma M-1 after one-time local irradiation of the tumour with 1000 R.