Structure-Function Analysis of NADE
Structure-Function Analysis of NADE
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DOI：
10.1074/jbc.m106342200
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发表时间：
2002-04
期刊：
影响因子：
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通讯作者：
J. Mukai;Shisako Shoji;M. Kimura;S. Okubo;H. Sano;P. Suvanto;Yin Li;S. Irie;Taka-Aki Sato
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文献类型：
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作者：
J. Mukai;Shisako Shoji;M. Kimura;S. Okubo;H. Sano;P. Suvanto;Yin Li;S. Irie;Taka-Aki Sato
神经生长因子 (NGF) 可通过激活低亲和力神经营养蛋白受体 p75NTR 诱导神经细胞凋亡。 NADE（p75NTR 相关细胞死亡执行蛋白）是一种 p75NTR 相关蛋白，通过与 293T、PC12 和 nnr5 细胞中 p75NTR 的死亡结构域相互作用，介导响应 NGF 的细胞凋亡（Mukai, J.、Hachiya, T.、Shoji-Hoshino, S.、Kimura, M. T.、Nadano, D.， Suvanto, P.、Hanaoka, T.、Li, Y.、Irie, S.、Greene, L. A. 和 Sato, T. A. (2000) J. Biol。化学。 275, 17566–17570）。我们对 NADE 进行了广泛的突变分析，以更好地表征其结构和功能特征。研究发现，截短一个最小区域（包括 NADE 的氨基酸残基 41-71）足以诱导细胞凋亡。指定的调节区域包括 C 端氨基酸残基 (72-112)，对于 NADE 诱导的细胞凋亡的 NGF 依赖性调节至关重要。此外，在富含亮氨酸的核输出信号（NES）序列（残基90-100）中进行氨基酸取代的突变体消除了NADE从细胞核到细胞质的输出。 NES 的突变还消除了 NADE 的自关联、其与 p75NTR 的相互作用以及 NGF 依赖性细胞凋亡。 NADE 片段（氨基酸残基 81-124）的表达阻断了少突胶质细胞中 NGF 诱导的细胞凋亡，表明该区域对 NGF/p75NTR 诱导的细胞凋亡具有显着的负面影响。这些研究确定了 NADE 的不同区域，这些区域参与调节 p75NTR 信号转导中的特定功能。
Nerve growth factor (NGF) can induce apoptosis in neural cells via activation of the low affinity neurotrophin receptor p75NTR. NADE (p75NTR-associated celldeath executor) is a p75NTR-associated protein that mediates apoptosis in response to NGF by interacting with the death domain of p75NTR in 293T, PC12, and nnr5 cells (Mukai, J., Hachiya, T., Shoji-Hoshino, S., Kimura, M. T., Nadano, D., Suvanto, P., Hanaoka, T., Li, Y., Irie, S., Greene, L. A., and Sato, T. A. (2000) J. Biol. Chem. 275, 17566–17570). We performed extensive mutational analysis on NADE, to better characterize its structural and functional features. Truncation of a minimal region, including amino acid residues 41–71 of NADE, was found to be sufficient to induce apoptosis. The designated regulatory region includes the C-terminal amino acid residues (72–112) and is essential for NGF-dependent regulation of NADE-induced apoptosis. Furthermore, the mutants with amino acid substitutions in the leucine-rich nuclear export signal (NES) sequence (residues 90–100) abolished the export of NADE from the nucleus to the cytoplasm. Mutation of the NES also abolished self-association of NADE, its interaction with p75NTR, and NGF-dependent apoptosis. Expression of a fragment of NADE (amino acid residues 81–124) blocked NGF-induced apoptosis in oligodendrocytes, suggesting that this region has a dominant negative effect on NGF/p75NTR-induced apoptosis. These studies identify distinct regions of NADE that are involved in regulating specific functions involved in p75NTR signal transduction.