Cyclobutane Pyrimidine Dimer Accumulation in Relation to UV-B Sensitivity in Rice Cultivars

Five rice ( Oryza sativa L.) cultivars, widely planted in South China, were grown in greenhouse with or without supplemental UV_B radiation at level of 13.6 kJ·m -2 ·d -1 . After 15 day_UV_B treatment, significant intraspecific differences were observed in plant height, photosynthetic rate and total biomass. Based on the total biomass accumulation, cultivar “Tesanai” was found to be the most sensitive, and cultivar “Luhuangzhan” was the most tolerant species to UV_B radiation. UV_B induced cyclobutane pyrimidine dimer (CPD) in rice DNA were quantified by ELISA with specific monoclonal antibody. CPD accumulations in DNA extracted from 5 rice cultivars were remarkably increased by UV_B radiation, and it was confirmed that there was a strong positive correlation between CPD accumulation and the inhibition of total biomass. Photorepair was proved to be the predominant mode of CPD repair in UV_B irradiated rice. Light_dependent removal of CPD was very fast as compared with dark repair. Different levels of CPD accumulation among rice cultivars were related with different capacity of CPD photorepair. Capacity of light_dependent CPD removal may play an important role in UV_B resistance of rice.

Molecular Dynamics Insights into Electron-Catalyzed Dissociation Repair of Cyclobutane Pyrimidine Dimer

Excess electrons are not only an important source of radiation damage,but also participate in the repair process of radiation damage such as cyclobutane pyrimidine dimer(CPD).Using ab initio molecular dynamics(AIMD)simulations,we reproduce the single excess electron stepwise catalytic CPD dissociation process in detail with an emphasis on the energy levels and molecular structure details associated with excess electrons.On the basis of the AIMD simulations on the CPD aqueous solution with two vertically added excess electrons,we exclude the early-proposed[2+2]-like concerted synchronous dissociation mechanism,and analyze the difference between the symmetry of the actual reaction and the symmetry of the frontier molecular orbitals which deeply impact the mechanism.Importantly,we propose a new model of the stepwise electron-catalyzed dissociation mechanism that conforms to the reality.This work not only provides dynamics insights into the excess electron catalyzed dissociation mechanism,but also reveals different roles of two excess electrons in two bond-cleavage steps(promoting versus inhibiting).

Effects of temperature on UV-B-induced DNA damage and photorepair in Arabidopsis thaliana

DNA damage in the form of cyclobutane pyrimidine dimers(CPDs) and (6-4) photoproducts(6-4PPs) induced by UV-B radiation in Arabidopsis thaliana at different temperatures was investigated using ELISA with specific monoclonal antibodies. CPDs and 6-4PPs increased during 3 h UV-B exposure, but further exposure led to decreases. Contrary to the commonly accepted view that DNA damage induced by UV-B radiation is temperature-independent because of its photochemical nature, we found UV-B-induction of CPDs and 6-4PPs in Arabidopsis to be slower at a low than at a high temperature. Photorepair of CPDs at 24℃ was much faster than that at 0℃ and 12℃, with 50% CPDs removal during 1 h exposure to white light. Photorepair of 6-4PPs at 12℃ was very slow as compared with that at 24℃, and almost no removal of 6-4PPs was detected after 4 h exposure to white light at 0℃. There was evidence to suggest that temperature-dependent DNA damage and photorepair could have important ecological implications.

Oxidative Splitting of a Pyrimidine Cyclobutane Dimer: A Pulse Radiolysis Study

The oxidative splitting process of cis-syn 1,3-dimethyluracil cyclobutane dimer (DMUD) in aqueous solution was investigated using pulse radiolysis technique. The results indicated that DMUD can be splitted into 1,3-dimethyluracil (DMU) by OH radicals (OH ·) and Br_2 radical anions (Br ·-_2), but not by azide radicals (N ·_3). The oxidative mechanisms that an H-abstracted from DMUD for OH · oxidative splitting and an electron transfer from DMUD to Br ·-_2, were suggested. Related kinetic parameters were determined.

Differences in dinucleotide frequencies of thermophilic genes encoding water soluble and membrane proteins

The occurrence frequencies of the dinucleotides of genes of three thermophilic and three mesophilic species from both archaea and eubacteria were investigated in this study. The genes encoding water soluble proteins were rich in the dinucleotides of purine dimers, whereas the genes encoding membrane proteins were rich in pyrimidine dimers. The dinucleotides of purine dimers are the counterparts of pyrimidine dimers in a double-stranded DNA. The purine/pyrimidine dimers were favored in the thermophiles but not in the mesophiles, based on comparisons of observed and expected frequencies. This finding is in agreement with our previous study which showed that purine/pyrimidine dimers are positive factors that increase the thermal stability of DNA. The dinucleotides AA, AG, and GA are components of the codons of charged residues of Glu, Asp, Lys, and Arg, and the dinucleotides TT, CT, and TC are components of the codons of hydrophobic residues of Leu, Ile, and Phe. This is consistent with the suitabilities of the different amino acid residues for water soluble and membrane proteins. Our analysis provides a picture of how thermophilic species produce water soluble and membrane proteins with distinctive characters: the genes encoding water soluble proteins use DNA sequences rich in purine dimers, and the genes encoding membrane proteins use DNA sequences rich in pyrimidine dimers on the opposite strand.

Protective role of electrophile-reactive glutathione for DNA damage repair inhibitory effect of dibromoacetonitrile

Dibromoacetonitrile(DBAN) is a disinfection byproduct(DBP) and linked with cancer in rodents, but the mechanism of its carcinogenicity has not been fully elucidated. We recently reported that DBAN induced inhibition of nucleotide excision repair(NER). In this study, we investigated if glutathione(GSH) is involved in the DBAN-induced inhibition of NER. Human keratinocytes Ha Ca T were pretreated with L-buthionine-(S,R)-sulfoximine(BSO) to deplete intracellular GSH. BSO treatment markedly potentiated the DBAN-induced NER inhibition as well as intracellular oxidation. The recruitment of NER proteins(transcription factor IIH, and xeroderma pigmentosum complementation group G) to DNA damage sites was inhibited by DBAN, which was further exacerbated by BSO treatment. Our results suggest that intracellular GSH protects cells from DBAN-induced genotoxicity including inhibition of DNA damage repair.