DNA damage caused by exposure to reactive oxygen species is one of the primary causes of DNA decay in most organisms. In plants, endogenous reactive oxygen species (ROS) are generated not only by respiration and photosynthesis, but also by active responses to certain environmental challenges, such as pathogen attack. Significant extracellular sources of activated oxygen include air pollutants such as ozone and oxidative effects of UV light and low-level ionizing radiation. Plants are well equipped to cope with oxidative damage to cellular macromolecules, including DNA. Oxidative attack on DNA generates both altered bases and damaged sugar residues that undergo fragmentation and lead to strand breaks. Recent advances in the study of DNA repair in higher plants show that they use mechanisms similar to those present in other eukaryotes to remove and/or tolerate oxidized bases and other oxidative DNA lesions. Therefore, plants represent a valuable model system for the study of DNA oxidative repair processes in eukaryotic cells.

Repair and tolerance of oxidative DNA damage in plants.

Gas chromatography-mass spectrometry with selected-ion monitoring was used to measure the yields of radiation-induced base products in aqueous solutions of native or heat-denatured DNA irradiated in the dose range 20-100 Gy. These DNA solutions were saturated with nitrous oxide, nitrogen, air or 20% oxygen in nitrous oxide during irradiation. The products measured were as follows: 5,6-dihydrothymine; 5-hydroxy-5,6-dihydrothymine; 5,6-dihydrothymine (thymine glycol); 5-hydroxy-5,6-dihydrocytosine; 5,6-dihydroxy-5,6- dihydrocytosine (cytosine glycol); 4,6-diamino-5-formamidopyrimidine; 7,8-dihydro-8-oxoadenine (8-hydroxyadenine); 2,6-diamino-4-hydroxy-5- formamidopyrimidine; and 7,8-dihydro-8-oxoguanine (8-hydroxyguanine). In oxygenated solutions, 5,6-dihydrothymine, 5-hydroxy-5,6-dihydrothymine and 5-hydroxy-5,6-dihydrocytosine were not formed. The yields of all products, other than 5,6-dihydrothymine, were greater in irradiated DNA samples from N2O-saturated solutions than from N2-saturated solutions. In N2-saturated solutions the yield of 8-hydroxyadenine was low and 8-hydroxyguanine was undetectable. Yields of pyrimidine products in heat-denatured DNA were greater than those in native DNA using all types of gases. However, the effects of DNA conformation on the yields of purine products were dependent on the type of gas used to saturate the irradiated DNA solutions. Yields of formamidopyrimidines were generally lower in solutions of DNA irradiated in the native than in the heat-denatured conformation. In air-saturated solutions of DNA, yields of 8-hydroxypurines were not influenced greatly by DNA conformation. In DNA solutions saturated with N2O/O2, 8-hydroxypurine formation was more favourable in the heat-denatured conformation than in the native conformation. On the other hand, in deoxygenated solutions, formation of 8-hydroxypurines was favoured in the native conformation. Data indicate that DNA conformation and the type of gas used to saturate the irradiated solutions have a profound influence on yields of base products in DNA.

Yields of radiation-induced base products in DNA: effects of DNA conformation and gassing conditions

The abasic site (apurinic/apyrimidinic site) is the most common lesion in DNA and is suggested to be an important intermediate in mutagenesis and carcinogenesis. We have recently reported a novel assay for the detection and quantitation of abasic sites in DNA [Kubo, K., Ide, H., Wallace, S. S., & Kow, Y. W. (1992) Biochemistry 31, 3703-3708]. In this assay, the aldehyde group in an abasic site is first modified by a probe bearing a biotin residue, called the Aldehyde Reactive Probe (ARP) and then the tagged biotin is quantified by an ELISA-like assay. However, in the previous study, ARP was prepared only in a crude form, and no solid chemical data concerning the structure and specificity of ARP were reported. In this study, an improved method for the preparative synthesis of ARP has been established, and its structure has been unambiguously characterized using spectroscopic means. In order to elucidate the specificity of ARP to DNA damages, ARP was incubated with a variety of damaged bases or nucleosides and the reaction mixtures were analyzed by HPLC. Of the 14 compounds tested for their reactivity to ARP, 2-deoxyribose (a model compound for an abasic site) and 5-formyluracil reacted with ARP. Interestingly, compounds bearing a formamide group such as formamidopyrimidine and deoxyribosylformamide did not react with ARP, indicating that ARP is specific to damages having an alkyl or allyl aldehyde group. Furthermore, the ability of ARP synthesized by the defined chemical route to detect abasic sites has been substantiated using natural DNA containing abasic sites. Potential applications and limitations of the ARP assay are discussed.

Synthesis and damage specificity of a novel probe for the detection of abasic sites in DNA

(1988). The metabolic activation of nitroheterocyclic therapeutic agents. Drug Metabolism Reviews: Vol. 19, No. 1, pp. 33-62.

The metabolic activation of nitroheterocyclic therapeutic agents.

The radiolysis of pyrimidines in aqueous solutions: an updating review.

SummaryThe effect of 20 electron-affinic compounds including nitroimidazoles, nitrofurans, nitrobenzenes, and quinones on the radiation-induced reaction of thymine in aqueous solution was studied under deaerated and N2O-saturated conditions. The radiolysis of thymine in aerated aqueous solution was also performed for comparison. Thymine decomposition was depressed to some extent by the addition of electron-affinic compounds in both deaerated and N2O-saturated solutions, while promoted in aerated solution. The radiolyses with varying concentration of misonidazole indicated that the depression of thymine decomposition can be attributed to a competition between thymine and electron-affinic compounds for the reactions with ·OH. Among the radiolysis products, the formation of thymine glycol was remarkably promoted by the addition of electron-affinic compounds. Irrespective of structures of the electron-affinic compounds, the G-value of thymine glycol increased in sigmoidal form with increasing one-electron red...

Radiation-induced hydroxylation of thymine promoted by electron-affinic compounds.

SummaryThe radiation-induced reduction of N1-alkyl substituted 2- and 5-nitroimidazoles in aqueous solution containing sodium formate or 2-propanol was studied at pH 7·0 ± 0·1 under deaerated conditions. Irrespective of 2- or 5-nitroimidazole, N1-unsubstituted nitroimidazoles (2-nitro- (1a), 4(5)-nitro- (2a), 2-methyl-4(5)-nitro (3a), and 4(5)-methyl-5(4)-nitro- (4a) imidazoles) were reduced stepwise to consume 6 electrons per molecule, whereas N1-alkyl substituted nitroimidazoles (1-methyl-2-nitro (5a) and 1-methyl-5-nitro (6a) imidazoles, misonidazole (7a), and metronidazole (8a)) reacted with consumption of 4 electrons. In accord with the stoichiometry for the reduction of N1-unsubstituted nitroimidazoles, the formation of the amino derivatives of (1a)–(4a) was shown by HPLC or colour identification tests. 4-Electron-reduction products of N1-alkyl substituted 2-nitroimidazoles (5a) and (7a) were characterized by 13C and 1H n.m.r. and FDMS measurements, indicating that the hydroxyamino derivative of (5a...

H. Ide

Papers

Radiation-induced hydroxylation of thymine promoted by electron-affinic compounds.

Radiation-induced Reduction of Nitroimidazole Derivatives in Aqueous Solution