• KSBB Journal
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• v.24 no.4
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• pp.403-409
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• 2009

H2AX, a crucial component of chromatin, is implicated in DNA repair, cell cycle check point and tumor suppression. The aim of this study was to identify direct binding partners of H2AX to regulate cellular responses to above mechanisms. Literature reviews and bioinformatical tools were attempted intensively to find binding partners of H2AX, which resulted in identifying two potential proteins, breast cancer-1 (BRCA1) and BRCA1-associated RING domain 1 (BARD1). Although it has been reported in vivo that BRCA1 co-localizes with H2AX at the site of DNA damage, their biochemical mechanism for H2AX were however only known that the complex monoubiquitinates histone monomers, including unphosphorylated H2AX in vitro. Therefore, it is important to know whether the complex directly interacts with H2AX, and also which regions of these are specifically mediated for the interaction. Using in vitro GST pull-down assay, we present here that BRCA1 and BARD1 directly bind to H2AX. Moreover, through combinational approaches of domain analysis, fragment clonings and in vitro binding assay, we revealed molecular details of the BRCA1-H2AX and BARD1-H2AX complex. These data provide the potential evidence that each of the BRCA1 nuclear localization signal (NLS) and BARD1 BRCA1 C-terminal (BRCT) repeat domain is the novel mediator of H2AX recognition.

• Journal of the Korean Society of Radiology
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• v.15 no.6
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• pp.781-788
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• 2021

Due to the high sensitivity to radiation, excessive exposure needs to be prevented by accurately measuring the dose irradiated to the skin during radiation therapy. Although clinical trials use dosimeters such as film, OSLD, TLD, glass dosimeter, etc. to measure skin dose, these dosimeters have difficulty in accurate dosimetry on skin curves. In this study, to solve these problems, we developed a skin dosimeter that can be attached according to human flexion and evaluated its response characteristics. For the manufacture of the dosimeter, lead oxide (PbO) with high atomic number (Z_Pb: 82, Z_O: 8) and density (9.53 g/cm³) and silicon binders that can bend according to human flexion were used. In the case of a dosimeter made of PbO material, the performance degradation has been prevented by using parylene and others due to the presence of degradation due to oxidation, but the previously used parylene is affected by bending, so a new form of passive layer was produced and applied to the skin dosimeter. The characteristic evaluation of the skin dosimeter was evaluated by analyzing SEM, reproducibility, and linearity. Through SEM analysis, bending was evaluated, reproducibility and linearity at 6 MeV energy were evaluated, and applicability was assessed with a skin dosimeter. As a result of observing the dosimeter surface through SEM analysis, the parylene passive layer PbO dosimeter with the positive layer raised to the parylene produced cracks on the surface when bent. On the other hand, no crack was observed in the silicon passive layer PbO dosimeter, which was raised to silicon passive layer. In the reproducibility measurement results, the RSD of the silicon passive layer PbO dosimeter was 1.47% which satisfied the evaluation criteria RSD 1.5% and the linearity evaluation results showed the R² value of 0.9990, which satisfied the evaluation criteria R² 9990. The silicon passive layer PbO dosimeter was evaluated to be applicable to skin dosimeters by demonstrating high signal stability, precision, and accuracy in reproducibility and linearity, without cracking due to bending.