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Subcellular localization of the proteins implicated in DNA damage-induced cell death

与 DNA 损伤诱导的细胞死亡有关的蛋白质的亚细胞定位

基本信息

批准号：
13670859
负责人：
MIYASHITA Toshiyuki
金额：
$ 1.86万
依托单位：
National Research Institute for Child Health and Development
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670859/
关键词：
apoptosis DNAdamage caspage Bim NF-κB isoform 細胞死 Fas p53 CNA損傷ミトコンドリア選択的スプライシング

项目摘要

Apoptosis or programmed cell death is a physiologic process that is regulated by evolutionarily conserved genes. Dysregulation of apoptosis can contribute to many major diseases including cancer, autoimmune disorders and certain neurodegenerative diseases. DNA damage-induced apoptosis is accomplished by proapoptotic members of the Bcl-2 family, such as Bim, and a family of cysteine proteases termed caspases. During the period of two years we got the results as follows.We have cloned and characterized six novel isoforms of human Bim, designated as Bimα1, α2, and β1-β4, which are generated by alternative splicing. Among the novel isoforms, only Bimα1 and α2 contained a BH3 domain and were proapoptotic, although less potent than the classical isoforms. These two isoforms localized, at least in part, in mitochondria. Expression profiles of bim isoforms were highly variable among normal tissues at least in humans, suggesting a tissue-specific transcriptional regulation of bim.Caspase-8 and -10 can induce NF-κB activation in protease-independent manner. Investigation of the signaling pathways leading to caspase-8 and -10-mediated NF-κB activation using the GST pull-down assay revealed that, among upstream kinases that activate NF-κB, NIK and RIP, but not RICK or IKKα/β could directly bind to caspase-8and -10. By using dominant-negative mutants and small interfering RNA technology, we also demonstrated that NIK and IKKα are required for this process.

细胞凋亡或程序性细胞死亡是一种由进化上保守的基因调控的生理过程。细胞凋亡的失调可导致许多主要疾病，包括癌症、自身免疫性疾病和某些神经退行性疾病。DNA损伤诱导的细胞凋亡是由Bcl-2家族的促细胞凋亡成员，如Bim和称为半胱氨酸天冬氨酸蛋白酶的半胱氨酸蛋白酶家族完成的。经过两年的研究，我们获得了以下结果：克隆并鉴定了6种新的人Bim异构体，分别命名为Bimα1、α2和β1-β4，它们是通过选择性剪接产生的。在新的亚型中，只有Bimα1和α2含有BH 3结构域，具有促凋亡作用，但作用不如经典亚型。这两种亚型至少部分位于线粒体中。Caspase-8和Caspase-10可诱导NF-κB的激活，但不依赖于蛋白酶。采用GST pull-down法研究caspase-8和caspase-10介导的NF-κB活化的信号通路发现，在激活NF-κB的上游激酶中，NIK和RIP可以直接与caspase-8和caspase-10结合，而RICK和IKKα/β不能。通过显性失活突变体和小干扰RNA技术，我们还证明了NIK和IKKα在这一过程中是必需的。