Role of p53 in radiation induced genomic instability

p53 在辐射诱导的基因组不稳定中的作用

• 批准号: 17201014
• 负责人: NIWA Ohtsura
• 金额: $ 27.04万
• 依托单位: National Institute of Radiological Sciences (2007)Kyoto University (2005-2006)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17201014/
• 关键词: p53; mouse pink-eyed unstable allele; homologous recombination; untargeted recombination; delayed recombination; Schizosaccharomvces pombe; DNA damage memory; p53; Sチェックポイント; 複製フォーク抑制; 相同組換え; 姉妹染色分体交換; ゲノム不安定性; 経世代影響; DNA複製フォーク進行; PCNA; プロ

We have previouslyreported the induction of mutations at the maternal alleles ofESTR locus and the pink-eyed unstable locus of Fl mice born to irradiated spermatozoa (PNAS 98, 1705, 2001; Radiat. Res. 157, 661, 2002)). These observations indicated the presence of cross-talk between X-irradiated paternal genome andunirradiated maternal genome whichinduces untargeted recombination in zygotic stage embryos and delayed recombination in fetuses. Further study has indicated that thiscross-talk is dependent on the function of p53 which mediates a novel p53 dependent S checkpoint in the zygotic stage mouse embryos (MCB, 22, 2220, 2002).In this study, we have first analyzed the p53 protein domain required for the novel p53 dependent S checkpoint Microinjection analyses of p53 protein with specific mutation demonstrated that this S checkpoint neither required transcription activation domain nor the ATM phosphorylation sites. However, the protein had mutation in the DNA binding domain was unable … More to carry out the S checkpoint function. These suggest that the p53 protein itself may bind to DNA to execute the S checkpoint (Oncogene24, 3229, 2005). In parallel with these results, DNA fiber analyses revealed that p53 dependent S checkpoint functioned in suppressing the progression of replication fork when cells were challenged by X-irradiationOOncogene25, 529, 2006).It can be envisaged that the slowing down of the replication fork progression may facilitate recombination between sisterchromatids. In order to test this possibility, primary mouse embryofibroblasts (MEFs) with the wild type p53 gene and those with the null allele were exposed to X-rays of up to 6 Gy. The frequency of SCE increased dose dependently in the wild type MEFs while that of the p53 null MEFs did not This links the p53 dependent S checkpoint to homologous recombination at least between sister chromatids.Our previous study indicated the increase in the frequency of recombinational mutation at the maternal pink-eyed unstable allele in the retinal pigment epithelial cells of sperm irradiated embryos. Since the retinal pigment epithelial cells start development at day 11 to day 12 of gestation, the frequency of recombination stays elevated for at least 11 to 12 days after fertilization with irradiated sperm. Indeed, the frequency of SCE was higher in primary MEFs of day 12 fetus fertilized by irradiated sperm (in preparation. Altogether, our 3 year mouse study has excavated a novel p53 dependent S checkpoint and its function toupregulate homologous recombination for at least 11 to 12 days in sperm irradiated embryos. In addition, they also demonstrated the DNA damage memory and its function in delayed recombination.In order to study the molecular mechanism, we have also tested Schizosaccharomyces pombe to see if the delayed and untargeted recombination can be induced. S. pombe exhibited upregulated recombination for around 10 cell cycle generations afterX-irradiation. The length of upregulated recombination was cell generation dependent rather than the absolute time dependent as shown by the temperature shift experiments. In addition, this upregulated recombination was not due to bystander factors since the phenomena were not affected by the presence of radical scavengers in the culture media. Concomitantly with the upregulation of the recombination frequency, Rad22 foci were observed for around 10 generations after irradiation. These studies also demonstrated the DNA damage memory and delayed recombination in S. pombe Less

我们以前已经报道了ESTR基因座的母亲等位基因的突变以及辐照精子出生的FL小鼠的粉红色眼睛不稳定基因座（PNAS 98，1705，2001; Radiat。Res。157，661，2002））。这些观察结果表明，X辐照的父亲基因组和未辐照的母体基因组之间存在串扰，这导致在二合期胚胎中未经靶向的重组和胎儿中的重组延迟。进一步的研究表明，这种交叉言论取决于p53的功能，p53介导了p53依赖的s检查点，在二元阶段小鼠的胚胎中介导了p53（MCB，22，2220，2002）。在这项研究中，我们首先分析了p53蛋白在p53依赖p53依赖的p53蛋白范围内均不相关的p53蛋白质，该p53依赖于p53依赖的p53蛋白p53蛋白p53蛋白p53蛋白p53蛋白酶均未进行过p53蛋白质的p53蛋白质。激活域或ATM磷酸化位点。然而，该蛋白在DNA结合结构域中具有突变是无法进行的……更多以执行S检查点功能。这些表明p53蛋白本身可以与DNA结合以执行S检查点（Oncogene24，3229，2005）。与这些结果同时，DNA纤维分析表明，当细胞受到X- iRradiationoonCogene25，529，529，2006的挑战时，依赖p53的检查点在抑制复制叉的进展中起作用。这可以预见，它可以预见，重复的fork进展可能会在姐妹中的重新构成sipterclycilsss s sipterclycins之间。为了测试这种可能性，用野生型p53基因和无效等位基因的初级小鼠胚胎纤维细胞（MEF）暴露于高达6 Gy的X射线。 The frequency of SCE increased dose dependently in the wild type MEFs while that of the p53 null MEFs did Not This links the p53 dependent S checkpoint to homologous recombination at least between sister chromatids.Our previous study indicated the increase in the frequency of recombinational mutation at the maternal pink-eyed unstable allele in the retinal pigment epithelial cells of sperm irradiated embryos.由于视网膜色素上皮细胞从妊娠的第11天到第12天开始发育，因此重组的频率在受精精子受精后至少保持升高11至12天。实际上，在第12天的主要MEF中，SCE的频率较高，这些胎儿被辐射的精子受精（在制备中。总的来说，我们的3年鼠标研究挖掘了一个新颖的p53依赖性p53检查点及其功能，其功能使同源性重组至少在精子辐射的胚胎中至少进行了11至12天的dna损坏，还显示了dna dne dne dere s dresem and dere dears Memory。机制，我们还测试了精神分裂症，以查看延迟和未靶向的重组。现象不受培养基中根治性的清道夫的影响。随着重组频率的上调，辐照后约10代，观察到Rad22灶。这些研究还证明了DNA损伤记忆和S. pombe S.

项目成果

Ku70/80 Modulates ATM and ATR Signaling Pathway in Response DNA Double Strand Breaks. [Poster presentation and also selected for workshop presentation

Ku70/80 在响应 DNA 双链断裂中调节 ATM 和 ATR 信号通路。

• 发表时间: 2007
• 作者: Tokoro T.;Watanabe A.;Kayanne H.;Nadaoka K.;Tamura H.;Nozaki K.;Kato K.;Negishi A.;丹羽 太貫;Igarashi,Y.;丹羽太貫;室崎 将史;丹羽太貫;Sato,E.;丹羽 太貫;Kuji,M.;栗政 明弘;Watanabe,K.;栗政 明弘
• 通讯作者: 栗政 明弘

Radiation induction of delayed recombination in S. pombe.

粟酒裂殖酵母延迟重组的辐射诱导。

• 期刊: DNA Repair (in press)
• 作者: Takada J;uematsu N;Shiraishi S;Toyoshima M;Matsumoto T;Niwa O.
• 通讯作者: Niwa O.

Radiation Research

辐射研究

• 发表时间: 2007
• 作者: Nakano M;Kodama Y;Ohtaki K;Niwa O;Nakamura N
• 通讯作者: Nakamura N

Indirect mechanisms of genomic instability and the biological significance of mutations at tandem repeat loci.

• DOI: 10.1016/j.mrfmmm.2006.01.015
• 发表时间: 2006-06
• 期刊: Mutation research
• 作者: O. Niwa

Radiation induction of delayed recombination in Schizosaccharomyces pombe

• DOI: 10.1016/j.dnarep.2008.04.006
• 发表时间: 2008-08-02
• 期刊: DNA REPAIR
• 影响因子: 3.8
• 作者: Takeda, Jun;Uematsu, Norio;Niwa, Ohtsura
• 通讯作者: Niwa, Ohtsura