Analysis of genes that are implicated in glucocorticoid-induced apoptosis

与糖皮质激素诱导的细胞凋亡有关的基因分析

• 批准号: 11670810
• 负责人: MIYASHITA Toshiyuki
• 金额: $ 2.3万
• 依托单位: National Children's Medical Research Center
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670810/
• 关键词: apoptosis; glucocorticoid; Bc1-2; caspase; Bax; Green Fluorescent protein; 細胞死; プロテアーゼ; 共焦点レーザー顕微鏡

Glucocorticoid (GC) is used for the treatment of leukemias or lymphomas because GC induces apoptosis in certain types of lymphocytes. However, the mechanism of how GC mediates apoptosis in unknown. In this research, we analyzed the subcellular localizations of some of the Bc1-2 family of proteins and caspases implicated in GC-induced apoptosis, using con focal microscopes, During the period of two years we got the results as follows.1. We have identified novel member of BH3-only Bcl-2 family member, Rad9, that binds to and colocalizes with Bc1-2 and promotes apoptosis.2. Since BH3 region of human and yeast Rad9 is conserved, it is speculated that unicellular organisms such as yeast may have prototypes of apoptosis-regulating genes.3. Bax is a proapoptotic member of Bcl-2 family, and the bax promoter contains four E-box sequences that are known to be potential c-myc binding sites. We have shown that c-myc can induce apoptosis through binding to the E-box elements ih bax promoter.4. Using green fluorescent protein as a tag, we have intensively analyzed subcellular localizations of eight apoptosis-related caspases. Most caspase members localized mainly in the cytoplasm. In contrast, caspase-2 was primarily a nuclear caspase. Remarkably, prodomain of caspase-8 and -10 formed striking filamentous structures that did not colocalize with any known organelles or fibers.

糖皮质激素（GC）用于治疗白血病或淋巴瘤，因为GC诱导某些类型的淋巴细胞凋亡。然而，GC介导细胞凋亡的机制尚不清楚。本研究利用共聚焦显微镜对GC诱导的细胞凋亡相关的Bcl-2家族蛋白和caspase进行了亚细胞定位分析，在为期两年的研究中，我们得到了以下结果.我们已经鉴定了BH 3-only Bcl-2家族成员的新成员Rad 9，其结合并共定位于Bc 1 -2并促进凋亡。由于人和酵母Rad 9的BH 3区域是保守的，推测单细胞生物如酵母可能具有凋亡调控基因的原型. Bax是Bcl-2家族中的一个促凋亡基因，其启动子区含有4个E-box序列，这些序列可能是c-myc的结合位点。我们已经证明c-myc可以通过与bax启动子中的E-box元件结合来诱导细胞凋亡.使用绿色荧光蛋白作为标签，我们已经深入分析了八个乳腺癌相关的半胱天冬酶的亚细胞定位。大多数caspase成员主要定位于细胞质中。相比之下，caspase-2主要是核caspase。值得注意的是，caspase-8和-10的前结构域形成了引人注目的丝状结构，不与任何已知的细胞器或纤维共定位。

项目成果

Miyashita, T, , et al.: "Expression of extended polyglutamine sequentially activates initiator and effector caspases."Biochem. Biophys. Res. Commun.. 257(3). 724-730 (1999)

Miyashita, T, 等人：“延长的聚谷氨酰胺的表达依次激活起始子和效应子半胱天冬酶。”Biochem。

Shikama, Y., et al.: "Comprehensive studies on subcellular localizations and cell death-inducing activities of eight GFP-tagged apoptosis-related caspases."Exp. Cell Res.. 264(2). 315-325 (2001)

Shikama, Y. 等人：“对八种 GFP 标记的细胞凋亡相关半胱天冬酶的亚细胞定位和细胞死亡诱导活性的综合研究。”

Shikama,Y., et al.: "Comprehensive studies on subcellular localizations and cell death-inducing activities of eight GFP-tagged apoptosis-related caspases."Experimental Cell Research. 264(2). 315-325 (2001)

Shikama,Y. 等人：“对八种 GFP 标记的凋亡相关半胱天冬酶的亚细胞定位和细胞死亡诱导活性的综合研究。”实验细胞研究。

Fujii, K., et al.: "γ-irradiation deregulates cell cycle controt and apoptosis in Devoid basal cell carcinoma syndrome-derived cells."Jpn. J.Cancer Res.. 90(12). 1351-1537 (1999)

Fujii，K.，等人：“γ-辐射解除了缺乏基底细胞癌综合征衍生细胞的细胞周期控制和细胞凋亡。”J.Cancer Res. 90(12)(1999)。

Okamura-Oho, Y., et al.: "Dentatorubral-pallidoluysian atrophy protein interacts through a proline-rich region near polyglutamine with the SH3 domain of an insulin receptor tyrosine kinase substrate"Hum. Mol. Genet.. 8(6). 947-957 (1999)

Okamura-Oho，Y.，等人：“齿状红斑-苍白球路易体萎缩蛋白通过聚谷氨酰胺附近富含脯氨酸的区域与胰岛素受体酪氨酸激酶底物的 SH3 结构域相互作用”Hum。