Analysis of a novel back-up repair system for UV-induced DNA damage in human cells.

分析一种新型后备修复系统，用于修复人体细胞中紫外线引起的 DNA 损伤。

• 批准号: 15310036
• 负责人: MATSUNAGA Tsukasa
• 金额: $ 7.55万
• 依托单位: Kanazawa University, Graduate School of Natural Science and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15310036/
• 关键词: DNA damage; nucleotide excision repair; ultraviolet light; cyclobutane pyrimidine dimer; (6-4) photoproduct; xeroderma pigmentosum; knock-out mouse; adaptive response; モノクローナル抗体

We have established a super-sensitive detection method for measuring cyclobutane pyrimidine dimers (CPDs) and (6-4)photoproducts (6-4PPs) induced by biological doses of ultraviolet (UV) light and suggested that human cells might have a novel repair system mainly for 6-4PPs other than nucleotide excision repair which has been thought to be only repair system for those lesions in humans so far. In this study, we have obtained the following findings.1.The removal of 6-4PPs was observed even in the embryonic fibroblasts derived from xpa(-/-) or xpg(-/-) knock-out mice (30-50% after 24 hr). In addition, the removal of CPDs was also detected in those cells with a less efficiency (20-30% after 48 hr).2.HeLa cells stably expressing AlwNI-type mutant XPA did not show any impairment of nucleotide excision repair activity, suggesting that this mutant XPA protein does not have a dominant-negative effect.3.We have further sensitized the detection method for CPDs, enabling us to determine the repair kinetics of CPDs in cells exposed to non-killing doses of Was low as 0.1 J/m^2. Under this condition, we have found that xeroderma pimentosum cells significantly remove CPDs (〜40% after 48hr).4.We could observe some weak enhancement of repair efficiency after UV irradiation of 1 J/m^2 when the human cells had been pie-exposed to 0.2 J/m^2. However, we need more extensive experiments to conclude whether the back-up repair system is under the control of adaptive responses.Taken together with those results, we concluded that mammalian cells have a certain back-up repair system, independent of XPA and XPG, which removes mainly 6-4PP but also CPDs with a less efficiency.

我们建立了一种检测生物剂量紫外线诱导的环丁烷嘧啶二聚体(CPDS)和(6-4)光产物(6-4pps)的超灵敏检测方法，提示人类细胞可能有一种新的修复系统，主要针对6-4pps，而不是核苷酸切除修复，这被认为是迄今为止人类对这些损伤唯一的修复系统。在本研究中，我们获得了以下发现：1.即使在xpa(-/-)或xpg(-/-)基因敲除小鼠的胚胎成纤维细胞中也观察到6-4pps的去除(24小时后30-50%)。2.稳定表达AlwNI型突变体XPA的HeLa细胞没有表现出核苷酸切除修复活性的任何损害，表明该突变的XPA蛋白不具有显性-负性效应。3.我们进一步敏化了CPDS的检测方法，使我们能够确定CPDS在低至0.1J/m^2的非致死剂量下的修复动力学。当人体细胞暴露在0.2J/m^2时，我们可以观察到1J/m^2的紫外线照射后修复效率有一些微弱的增强。然而，我们还需要更广泛的实验来判断后备修复系统是否受适应性反应的控制。综合这些结果，我们得出结论，哺乳动物细胞有一定的后备修复系统，它不依赖于XPA和XPG，主要清除6-4PP，但也有效率较低的CPD。

项目成果

Identification of the XPG Region That Causes the Onset of Cockayne Syndrome by Using Xpg Mutant Mice Generated by the cDNA-Mediated Knock-In Method

• DOI: 10.1128/mcb.24.9.3712-3719.2004
• 发表时间: 2004-05
• 期刊: Molecular and Cellular Biology
• 影响因子: 5.3
• 作者: N. Shiomi;S. Kito;Masaki Oyama;T. Matsunaga;Y. Harada;M. Ikawa;M. Okabe;T. Shiomi

Lan, L.: "Functional and physical interactions between ERCC1 and MSH2 for resistance to cis-platinum in mammalian cells"DNA Repair. 3(2). 135-143 (2004)

Lan, L.：“ERCC1 和 MSH2 之间的功能和物理相互作用对哺乳动物细胞中顺铂的抵抗力”DNA 修复。

UV-B protective effect of a polyacylated anthocyanin, HBA, in flower petals of the blue morning glory, Ipomoea tricolor cv. Heavenly Blue

• DOI: 10.1016/j.bmc.2005.01.011
• 发表时间: 2005-03-15
• 期刊: BIOORGANIC & MEDICINAL CHEMISTRY
• 影响因子: 3.5
• 作者: Mori, M;Yoshida, K;Kondo, T
• 通讯作者: Kondo, T

cDNA cloning of the chicken DDB1 gene encoding the p127 subunit of damaged DNA-binding protein.

编码受损 DNA 结合蛋白 p127 亚基的鸡 DDB1 基因的 cDNA 克隆。

• 发表时间: 2003
• 期刊: Gene Genet.Syst. 78(2)
• 作者: Fu;D.
• 通讯作者: D.

Disruption of the AtREV3 gene causes hypersensitivity to ultraviolet B light and γ-rays in Arabidopsis inplication of the presence of a translesion synthesis mechanism in plants.

AtREV3 基因的破坏会导致拟南芥对紫外线 B 光和 γ 射线过敏，这表明植物中存在跨损伤合成机制。

• 发表时间: 2003
• 期刊: Plant Cell 15
• 作者: Sakamoto;A.;Lan;V.T.T.;Hase;Y.;Shikazono;N.;Matsunaga;T.;Tanaka;A.
• 通讯作者: A.