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.

我们试图建立从全身到分子水平的氚水生物剂量测量系统。1)建立对辐射敏感的Rev1转基因小鼠。我们鉴定了UMUC/DinB/XPV基因家族中的人和小鼠的Rev1基因，该基因在自发突变中起重要作用。对小鼠Rev1蛋白的生化分析表明，小鼠Rev1蛋白具有与人Rev1蛋白相同的脱氧胞苷转移酶活性。辐射诱导小鼠REV1在胚胎成纤维细胞中表达。在此基础上，我们尝试建立Rev1转基因小鼠，以建立辐射超敏小鼠模型。我们将金属蛋白启动子下游携带mRev1的MT-1载体导入C57BL小鼠体内。我们已经有36只小鼠携带mRevl。我们将检查mRev1的表达。2)氚水分子生物剂量学系统的建立在辐射损伤DNA双链断裂的修复机制中，历史型的H_2AX、Ku70、Ku80和Tip60被认为起着重要的作用。我们观察到照射后H_2AX被磷酸化(γ-H_2AX)并形成病灶。为了将焦点形成用于DNA可加成链断裂的剂量测定，我们纯化了组成H_2AX的复合体，并通过MS/MS光谱分析鉴定了它的新组分。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，以检测DNAdouble链断裂处的病灶。此外，我们还发现，辐射后NBS1定位于γ-H_2AX焦点。病灶数目和DNA双链断裂之间的关系正在调查中。

项目成果

Sugaya, Y.: "Hyper-processive and slower DNA chain elongation catalysed by DNA polymerase III holoenzyme purified from the dnaE173 mutator mutant of Escherichia coli"Genes Cells. 7. 151-156 (2002)

Sugaya, Y.：“从大肠杆菌 dnaE173 突变突变体中纯化的 DNA 聚合酶 III 全酶催化的超持续且较慢的 DNA 链延伸”Genes Cells。

Tauchi, H.: "Nijmegenbreakage syndrome gene, NBS1,and moleculer links to factors for genome stability"Oncogene. 21. 8967-8980 (2002)

Tauchi, H.：“奈梅亨断裂综合征基因、NBS1 和分子与基因组稳定性因素的联系”癌基因。

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)

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。