Analysis of mutations induced in cells derived from DNA repair-deficient hereditary disease patients.

DNA 修复缺陷型遗传性疾病患者细胞中诱导的突变分析。

• 批准号: 02671046
• 负责人: YAGI Takashi
• 金额: $ 1.34万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02671046/
• 关键词: DNA repair; Mutation; xeroderma pigmentosum; Ataxia telangectasia; Cancer-prone disease; Ultraviolet light; Gamma ray

To assess the contribution to mutagenesis by DAN repair defects, mutagen-treated shuttle vector plasmids were passed through fibroblasts derived from xeroderma pigmentosum(XP)patients in 3 different DNA repair complementation groups(A, C and F), ataxia telangectasia(AT)patients and mice.In comparison to DNA repair proficient normal human cells, the UV-treated plasmed passed through the XP-A, C or F cells showed fewer surviving plasmids(XP-A less than C and F)and a higher frequency of mutated plasmids(XP-A greater than C and F). Among base substitution mutations, the major type of the substitution was G : C -> A : T transition in all 3 cell lines. The XP-A, C and F cells revealed a hlgher frequency of G : C -> A : T transition along with a lower frequency of transverslons compared to the normal line. The spectrum of mutations In the XP-A cells was similar to that in the XP-C and F cells. Most single base substitution mutations occurred at G : C base pairs in which the 5'-nelghboring base of the cytosine was a pyrimidine. In spontaneous mutants, more than 50% mutants have a deletion, and the rest have base substitutions with a high frequency of transversions. Among UV-Induced base substitution mutants from mouse cells, 91% mutants have G : C -> A : T transition. gamma -ray-irradiated plasmids passed through AT and normal cells showed no significant difference in the survival and a frequency of mutated plasmed. Efficiency of rejoining of vector DNA restricted by an endonuclease was similar between AT and normal cells, but fidelity of the rejoining is higher in the normal than AT cells. A frequency of methyl nitrosourea-induced mutations, especially G : C -> A : T transition was higher in O^6-alkylguanine-repair deficient mouse cells than the proficient mouse cells.In this study, a role of DNA repair for the Induction of mutation in molecular level was clarified by using shuttle vectors and DNA repair deficient cells.

为了评估 DAN 修复缺陷对诱变的贡献，将诱变剂处理的穿梭载体质粒穿过来自 3 个不同 DNA 修复互补组（A、C 和 F）的色素性干皮病 (XP) 患者、共济失调性毛细血管扩张 (AT) 患者和小鼠的成纤维细胞。与 DNA 修复能力良好的正常人类细胞相比，经紫外线处理的质粒通过了 DAN 修复缺陷对诱变的贡献。 XP-A、C或F细胞显示出较少的存活质粒（XP-A小于C和F）和较高的突变质粒频率（XP-A大于C和F）。在碱基取代突变中，所有 3 个细胞系中的主要取代类型是 G : C -> A : T 转变。与正常细胞系相比，XP-A、C 和 F 细胞显示出更高的 G : C -> A : T 转变频率以及更低的横向频率。 XP-A 细胞中的突变谱与 XP-C 和 F 细胞中的突变谱相似。大多数单碱基取代突变发生在 G : C 碱基对上，其中胞嘧啶的 5'-邻近碱基是嘧啶。在自发突变体中，超过50%的突变体存在缺失，其余突变体存在碱基取代，且颠换频率较高。在小鼠细胞的紫外线诱导碱基取代突变体中，91% 的突变体具有 G : C -> A : T 转变。伽马射线照射的质粒通过AT和正常细胞后，其存活率和突变质粒的频率没有显着差异。 AT和正常细胞之间受核酸内切酶限制的载体DNA重新连接的效率相似，但正常细胞中重新连接的保真度高于AT细胞。 O^6-烷基鸟嘌呤修复缺陷小鼠细胞中甲基亚硝基脲诱导的突变频率，特别是 G : C -> A : T 转变频率高于正常小鼠细胞。 在本研究中，通过使用穿梭载体和 DNA 修复缺陷细胞，阐明了 DNA 修复在分子水平上诱导突变的作用。

项目成果

Wang. S., Nishigori, C., Yagi, T. and Takebe, H.: "Reduced DNA repair in progeria cells and effects of gamma -ray irradiation on UV-induced unscheduled DNA synthesis in normal and progeria cells." Mutation Research. 256. 59-66 (1991)

王.

Yagi, T., Sato, M., Tatsumi-Miyajima, J. and Takebe, H.: "UV-induced base substitution mutations in a shuttle vector plasmid propagated in group C xeroderma pigmentosum cells." Mutation Research. (1992)

Yagi, T.、Sato, M.、Tatsumi-Miyajima, J. 和 Takebe, H.：“在 C 组色素性干皮病细胞中繁殖的穿梭载体质粒中紫外线诱导的碱基取代突变。”

Takashi Yagi: "Analysis of point mutations in an ultravioletーirradiated shuttle vector plasmid propagated in cells from Japanese xeroderma pigmentosum patients in complementation groups A and F." Cancer Research. 51. 3177-3182 (1991)

Takashi Yagi：“在互补组 A 和 F 的日本色素性干皮病患者细胞中繁殖的紫外线照射穿梭载体质粒的点突变分析”，51。 3177-3182 (1991)。

Mayumi Sato: "Protective effects of sodium selenite on killing and mutation by N-methyl-N^1-nitro-N-nitrosoguanidine in E.coli." Mutation Research. 250. 73-77 (1991)

Mayumi Sato：“亚硒酸​​钠对大肠杆菌中 N-甲基-N^1-硝基-N-亚硝基胍的杀灭和突变的保护作用。”

Shin-ichi Moriwaki: "Analysis of N-methyl-N-nitrosourea-induced mutations in a shuttle vector plasmid propagated in mouse O^6-methylguanine-DNA methyltransferase-deficient cells in comparison with proficient cells." Cancer Research. 51. 6219-6223 (1991)

Shin-ichi Moriwaki：“与正常细胞相比，在小鼠 O^6-甲基鸟嘌呤-DNA 甲基转移酶缺陷细胞中繁殖的穿梭载体质粒中 N-甲基-N-亚硝基脲诱导的突变分析。”