Development of molecular bio-dosimeiry and monitor mice for the detection of radiation dose exposed by tritium water

开发用于检测氚水辐射剂量的分子生物剂量学和监测小鼠

• 批准号: 12558049
• 负责人: KAMIYA Kenji
• 金额: $ 7.3万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12558049/
• 关键词: Rev1; tritium; translesion DNA repair; DNA repair; H2AX; bio-dosimetry; radiation; DNA double strand break; Revl; 遺伝子損傷; 2重鎖切断; 自然突発異変; 誤りがちなDNA合成; translesion; トリチウム水; 突然変異; トランスジェニックマウス; 損傷乗り越えDNA複製

We tried to develop bio-dosimetry system of tritium water at the level from whole body to molecule.1) Development of Revl transgenic mice which are sensitive to radiation exposureWe characterized mouse and human Revl gene which was a member of the UmuC/DinB/XPV gene family, played important roles in spontaneous mutations. Biochemical analysis of the mouse Revl protein revealed that the mouse Revl protein possessed a deoxycytidyl transferase activity as human REV1 protein. The expression of mouse Revl in embryonic fibroblasts was induced by radiation exposure. Based on these knowledge, we try to establish Revl transgenic mouse to develop the hypersensitive mouse model to radiation exposure. We introduced MT-1 plasmid carrying mRevl down stream the metallotuionein promoter into C57BLmice. We already got 36 mice carrying mRevl. We will check the expression of mRevl.2) Development of molecular bio-dosimetry system for tritium waterIn repair machinery of DNA double trand break by radiation, Historic H2AX, Ku70, Ku80 and Tip60 are thought to play an important role. We observed that H2AX was phosphorylated (γ-H2AX) and formed foci after irradiation. In order to use the foci formation for the dosimetry of DNAdouble strand breaks, we purify the constitutive H2AX complex and identify its new components by MS/MS spectrometric analysis. We prepared antibody against these components and performed the immunohistochemical analysis to detect the foci at DNAdouble strand breaks. Furthermore, we found that NBS1 localized toγ-H2AX foci after radiation exposure. Relationship between numbers of foci and DNAdouble strand breaks is under investigation.

我们尝试开发从全身到分子水平的氚水生物剂量测定系统。1）对辐射敏感的Revl转基因小鼠的开发我们对小鼠和人类Revl基因进行了表征，该基因是UmuC/DinB/XPV的成员基因家族，在小鼠 Revl 蛋白的自发突变中发挥重要作用，小鼠 Revl 蛋白具有与人 REV1 蛋白一样的脱氧胞苷转移酶活性。基于这些知识，我们尝试建立Revl转基因小鼠以开发对辐射暴露过敏的小鼠模型，我们将携带mRevl下游的MT-1质粒导入C57BL小鼠中。携带mRevl的小鼠我们将检查mRevl的表达。2)氚水分子生物剂量测定系统的开发DNA双链修复机制由于辐射断裂，Historic H2AX、Ku70、Ku80 和 Tip60 被认为起着重要作用。我们观察到 H2AX 被磷酸化（γ-H2AX）并在辐射后形成焦点，以便将焦点形成用于 DNA 双链的剂量测定。断裂后，我们纯化了组成型 H2AX 复合物，并通过 MS/MS 光谱分析鉴定了其新成分，我们制备了针对这些成分的抗体并进行了免疫组织化学分析。检测DNA双链断裂处的焦点此外，我们发现放射暴露后NBS1定位于γ-H2AX焦点，并且正在研究焦点数量与DNA双链断裂之间的关系。

项目成果

Ikura, T.: "Chromatin dynamics and DNA repair"Front. Biosci.. 8. 149-155 (2003)

Ikura, T.：“染色质动力学和 DNA 修复”前面。

Fukuda. S.: "Murine and human SDF2L1 is an endoplasmic reticulum stress-inducible gene and encodes a new member of the Pmt/rt protein family."Biochem. Biophys. Res. Commun.. 280(1). 407-414 (2001)

福田。

Nitta, Y.: "Effects of radioactive iodine (131I) on the thyroid of newborn, pubertal and adult rats."Proceedings of International Symposium on Radiation and Homeostasis (Ed. by Sugahara, T.), Elsevier, Amsterdam, Netherland. 127-131 (2002)

Nitta, Y.：“放射性碘 (131I) 对新生、青春期和成年大鼠甲状腺的影响。”国际辐射与稳态研讨会论文集（由 Sugahara, T. 编辑），爱思唯尔，阿姆斯特丹，荷兰。

J. Kobayashi: "NBS1 localizes to γ-H2AX foci through interaction with the FHA/BRCT domain."Curr. Biol.. 12. 1846-1851 (2002)

J. Kobayashi：“NBS1 通过与 FHA/BRCT 结构域相互作用定位于 γ-H2AX 焦点。”Curr.12. 1846-1851 (2002)

Masuda, Y: "Deoxycytidyl transferase activity of the human REV1 protein is closely associated with the conserved polymerase domain"J. Biol. Chem.. 276. 15051-15058 (2001)

Masuda, Y：“人 REV1 蛋白的脱氧胞苷基转移酶活性与保守的聚合酶结构域密切相关”J.