UVA1 genotoxicity is mediated not by oxidative damage but by cyclobutane pyrimidine dimers in normal mouse skin

UVA1 induces the formation of 8-hydroxy-2′-deoxyguanosines (8-OH-dGs) and cyclobutane pyrimidine dimers (CPDs) in the cellular genome. However, the relative contribution of each type of damage to the in vivo genotoxicity of UVA1 has not been clarified. We irradiated living mouse skin with 364-nm UVA1 laser light and analyzed the DNA damage formation and mutation induction in the epidermis and dermis. Although dose-dependent increases were observed for both 8-OH-dG and CPD, the mutation induction in the skin was found to result specifically from the CPD formation, based on the induced mutation spectra in the skin genome: the dominance of C → T transition at a dipyrimidine site. Moreover, these UV-specific mutations occurred preferentially at the 5′-TCG-3′ sequence, suggesting that CpG methylation and photosensitization-mediated triplet energy transfer to thymine contribute to the CPD-mediated UVA1 genotoxicity. Thus, it is the CPD formation, not the oxidative stress, that effectively brings about the genotoxicity in normal skin after UVA1 exposure. We also found differences in the responses to the UVA1 genotoxicity between the epidermis and the dermis: the mutation induction after UVA1 irradiation was suppressed in the dermis at all levels of irradiance examined, whereas it leveled off from a certain high irradiance in the epidermis

Biological effects of heavy ion near the Bragg peak

10th International Workshop on Radiation Damagea to DN

Biological effects of heavy ion near the Bragg peak

10th International Workshop on Radiation Damagea to DN

DNA damage induced by low energy ions near the Bragg peak

Molecular Imaging of the Process of DNA Damage Induction and It\u27s Repai

The involvement of cell cycle checkpoint-mutations in the mutagenesis induced in Drosophila by a longer wavelength light band of solar UV

Solar ultraviolet radiation is considered to be injurious rather than necessary for most organisms living on the earth. It is reported that the risk of skin cancer in humans increased by the depletion of the ozone layer. We have examined the genotoxicity of solar ultraviolet, especially the longer wavelengths light, using Drosophila. Recently, we have demonstrated that light of wavelengths up to 340 nm is mutagenic on Drosophila larvae. Using an excision repair-deficient Drosophila strain (mus201), we have obtained results suggesting that the lesion caused in larvae by the 320 nm-light irradiation may be similar to the damage induced by irradiation at 310 nm, and that light of 330 and 340 nm may induced damage different from that induced by 310 and 320 nm-light. To examine the difference in DNA damage induced by light of particular wavelength, we performed monochromatic irradiation on larvae of two Drosophila strains; one excision repair-deficient (mei-9) and another postreplication repair-deficient (mei-41). 310 and 320 nm-light was more mutagenic in the mei-9 strain than mei-41, whereas 330 and 340 nm-light was more mutagenic in mei-41 than in mei-9. It is demonstrated that the mei-41 gene is a homologue of the human atm gene which is responsible for a cell cycle checkpoint. This result suggests that 310-320 nm-light induces DNA damage that is subject to nucleotide excision repair (NER) and that 3300-360 nm-light causes damage to be recognized by the cell cycle checkpoint but it is not repairable by NER

Visualization of Heavy Ion Tracks by Labeling 3\u27-OH Termini of Induced DNA Strand Breaks

African green monkey kidney cells, CV-1, were irradiated with Carbon ions (LET: 735 keV/um Argon ions (LET: 3,000 keV/um) to visualize ion tracks through the cell nucleus by labeling the 3\u27-OH termini result of DNA strand breaks. The 3\u27-OH termini of DNA were labeled with BrdU-triphosphate catalyzed by TdT. This method of TUNEL (TdT-mediated dUTP Nick End labeling) is based on the specific binding of TdT to 3\u27-OH termini of DNA. Subsequent immuno-fluorescent staining with the primary monoclonal antibody against BrdU, followed by a secondary antibody of Alexa Fluor 488, was performed to visualize the BrdU labeled DNA termini. Images of the cell nuclei were acquired by confocal laser microscopy. When cell monolayers were irradiated perpendicularly with argon ions, induced DSBs in cell nuclei were identifiable as fluorescent spots. In another irradiation setup, when cells were irradiated at a small angle with incident argon ions, DNA strand breaks were detected as fluorescent stripes across the cell nucleus. These results demonstrate the induction of 3\u27-OH termini at sites of DNA strand breaks along Argon ion tracks

Enhanced Cell Inactivation and Double-strand Break Induction in V79 Chinese Hamster Cells by Monochromatic X-rays at Phosphorus K-shell Absorption Peak

To investigate an enhancement of cell inactivation and DNA double-strand break (DSB) induction by the K-shell ionization of phosphorus atoms and Auger electrons, monochromatic X-rays on and below the phosphorus K-shell absorption peak, 2.153 keV and 2.147 keV were exposed to Chinese hamster lung fibroblast V79 cells. Survival fractions were plotted against exposure, Ψ [nC/kg] and the linear-quadratic (LQ) model was adapted to estimate the parameters, α and β, of the survival curves. DSB induction rate [DSB/cell/Ψ] was estimated from the measured fractions of induced DNA fragments below 4.6 Mbp (Fk<4.6Mbp), which were determined using pulse field gel electrophoresis. As results, both cell inactivation and DSB induction rate of on the peak were significantly higher compared to that of the below. However, when converting Ψ to absorbed dose (Gy) of cell nucleus, the enhanced effect was only observed for parameter α, and not for a survival dose (Gy) of 37%, 10%, and 1% nor for a DSB induction rate. Our findings indicate that enhancement of cell inactivation and DSB induction were due to the additional dose delivered to the DNA and more complex DSB lesions were induced due to the release of phosphorus K-shell photoelectrons and Auger electrons