Identification of molecular mechanisms of neuronal cell death through ASK1

通过ASK1鉴定神经细胞死亡的分子机制

• 批准号: 13557156
• 负责人: NISHITOH Hideki
• 金额: $ 9.02万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 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/en/grant/KAKENHI-PROJECT-13557156/
• 关键词: Apoptosis; MAP kinase; ASK1; 細胞死; 神経変性疾患; 機能回復

In this research project, we have shown the following three results.1) Apoptosis Signal-regulating Kinase (ASK) 1 is activated in response to various cytotoxic stresses including TNF, Fas and reactive oxygen species (ROS) such as H2O2 and activates c-Jun NH2-terminal Kinase (JNK) and p38. We have shown by deleting ASK1 in mice that TNF- and H2O2-induced sustained activations of JNK and p38 are lost in ASK1V- embryonic fibroblasts, and that ASK1-/- cells are resistant to TNF- and H2O2-induced apoptosis. TNTMnduced apoptosis requires ROS-dependent activation of ASKl-JNK/p38 pathways. Thus, ASK1 is selectively required for TNF- and oxidative stress-induced sustained activations of JNK/p38 and apoptosis. (EMBO Reports, Vol. 2, p222-228 2001)2) A yeast two-hybrid screening identified a serine/threonine protein phosphatase 5 (PP5) as a binding partner of ASK1. PP5 directly dephosphorylated an essential phospho-threonine residue within the kinase domain of ASK1 and thereby inactivated ASK1 ac … More tivity in vitro and in vivo. The interaction between PP5 and ASK1 was induced by H2O2 treatment and was followed by the decrease in ASK1 activity. PP5 inhibited not only H2O2-induced sustained activation of ASK1 but also ASK 1-dependent apoptosis. Thus, PP5 appears to act as a physiological inhibitor of ASKl-JNK/p38 pathways by negative feedback. (EMBO Journal, Vol. 20, p6028-<3036,2001)3) Accumulation of misfolded proteins within the ER lumen induces cellular stress and cell death, and ER stress has been implicated in human neurodegenerative disorders. However, the molecular mechanism of ER stress-induced cell death was controversial. We have shown the role of ASK1 in the ER stress signaling. Activated IRE1, ER-resident type I transmembrane serine/threonine protein kinase, recruits TRAF2 and ASK1. In untransfected cells, ASK1 is activated by the ER stressors. By using ASK1V-cells, ASK1 was shown to be required for the ER stress-induced JNK activation and apoptosis. These results indicate that IRE1-TRAF2-ASK1 axis is essential for the ER stress-induced JNK activation and apoptosis. Furthermore, we have shown the evidence that this ER stress-induced cell death pathway plays a central role in the pathogenesis of neurodegenerative disorders. (Genes & Development Vol. 16, p!345-1355, 2002) Thus, ER stress-mediated ASK1 pathway may be one of the therapuitic targets for diseases. Less

在本研究项目中，我们得到了以下三个结果：1）细胞凋亡信号调节激酶（ASK）1在各种细胞毒性应激（包括TNF、Fas和活性氧（ROS）如H2 O2）中被激活，并激活c-Jun NH 2-末端激酶（JNK）和p38。我们已经证明，通过在小鼠中删除ASK 1，TNF-和H2 O2-诱导的JNK和p38的持续激活在ASK 1V-胚胎成纤维细胞中丢失，并且ASK 1-/-细胞对TNF-和H2 O2-诱导的凋亡具有抗性。TNF诱导的细胞凋亡需要ASK 1-JNK/p38途径的ROS依赖性活化。因此，ASK 1是选择性地需要TNF-和氧化应激诱导的持续激活JNK/p38和凋亡。(EMBO 2）酵母双杂交筛选鉴定了丝氨酸/苏氨酸蛋白磷酸酶5（PP 5）作为ASK 1的结合配偶体。PP 5直接使ASK 1激酶结构域中的一个必需的磷酸化苏氨酸残基去磷酸化，从而使ASK 1失活。 ...更多信息 体外和体内的活性。H_2O_2处理诱导了PP_5与ASK_1之间的相互作用，随后ASK_1活性下降。PP 5不仅抑制H2 O2诱导的ASK 1的持续激活，而且抑制ASK 1依赖的凋亡。因此，PP 5似乎通过负反馈充当ASK 1-JNK/p38途径的生理抑制剂。(EMBO Journal，Vol.20，p6028-<3036，2001）3）错误折叠的蛋白质在ER腔内的积累诱导细胞应激和细胞死亡，并且ER应激与人类神经变性疾病有关。然而，ER应激诱导细胞死亡的分子机制仍存在争议。我们已经显示了ASK 1在ER应激信号传导中的作用。激活的IRE 1，ER驻留I型跨膜丝氨酸/苏氨酸蛋白激酶，招募TRAF 2和ASK 1。在未转染的细胞中，ASK 1被ER应激物激活。通过使用ASK 1V-细胞，ASK 1被证明是ER应激诱导的JNK激活和凋亡所必需的。这些结果表明，IRE 1-TRAF 2-ASK 1轴在ER应激诱导的JNK激活和凋亡中是必需的。此外，我们已经证明了这种ER应激诱导的细胞死亡途径在神经退行性疾病的发病机制中起着重要作用。（Genes & Development Vol. 16，p！345-1355，2002）因此，ER应激介导的ASK 1途径可能是疾病的治疗靶标之一。少

项目成果

Matsuura, H. et al.: "Phosphorylation-dependent scaffolding role of JSAP1/JIP3 in the ASK 1-INK signaling pathway: a new mode of regulation of the MAP kinase cascade"J.Biol. Chem.. 277. 40703-40709 (2002)

Matsuura, H. 等人：“ASK 1-INK 信号通路中 JSAP1/JIP3 的磷酸化依赖性支架作用：MAP 激酶级联调节的新模式”J.Biol。

Saeki, K: "Oxidation-triggered c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein (MAP) kinase pathways for apoptosis in human leukaemic cells stimulated by epigallocatechin-3-gallate (EGCG) : a distinct pathway from those of chemically induc

Saeki, K：“表没食子儿茶素-3-没食子酸酯 (EGCG) 刺激的人白血病细胞中，氧化触发的 c-Jun N 末端激酶 (JNK) 和 p38 丝裂原激活蛋白 (MAP) 激酶通路导致细胞凋亡：这是一条与

Nishitoh, H., et al.: "ASK1 is essential for endoplasmic reticulum stress-induced neuronal cell death triggered by expanded polyglutamine repeats."Genes & Dev.. 16. 1345-1355 (2002)

Nishitoh, H. 等人：“ASK1 对于由扩展的多聚谷氨酰胺重复序列引发的内质网应激诱导的神经元细胞死亡至关重要。”

Saeki K. et al.: "Oxidation-Triggered c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein (MAP) kinase pathways for apoptosis in human leukaemic cells stimulated by epigallocatechin-3-gallate (EGCG) : a distinct pathway from those ofchemically

Saeki K. 等人：“表没食子儿茶素-3-没食子酸酯 (EGCG) 刺激的人白血病细胞中氧化触发的 c-Jun N 末端激酶 (JNK) 和 p38 丝裂原激活蛋白 (MAP) 激酶通路导致细胞凋亡：a

Matsuura, H. et al.: "Phosphorylation-dependent scaffolding role of JSAP1/JIP3 in the ASK1-JNK signaling pathway : a new mode of regulation of the MAP kinase cascade"J. Biol. Chem.. 277. 40703-40709 (2002)

Matsuura, H. 等人：“ASK1-JNK 信号通路中 JSAP1/JIP3 的磷酸化依赖性支架作用：MAP 激酶级联调节的新模式”J.