F1Aα, a death receptor-binding protein homologous to the Caenorhabditis elegans sex-determining protein, FEM-1, is a caspase substrate that mediates apoptosis
F1Aα, a death receptor-binding protein homologous to the Caenorhabditis elegans sex-determining protein, FEM-1, is a caspase substrate that mediates apoptosis
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DOI：
10.1074/jbc.274.45.32461
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发表时间：
1999-11-05
影响因子：
4.8
通讯作者：
Yu, VC
中科院分区：
文献类型：
--
作者：
Chan, SL;Tan, KO;Yu, VC
细胞凋亡是一个进化上保守的过程，在基本上所有的多细胞生物中，它对组织的稳态和发育包括性别决定都是至关重要的。在这里，我们报告的克隆锚蛋白重复含有蛋白质，称为F1A α，在酵母双杂交筛选使用Fas（CD95/APO-1）的胞质结构域作为诱饵。氨基酸序列分析表明，F1 A alpha与秀丽隐杆线虫的性别决定蛋白FEM-1具有广泛的同源性，这是雄性表型各个方面发育所必需的。F1A α与Fas和肿瘤坏死因子受体1的胞质结构域相关，Fas和肿瘤坏死因子受体1是"死亡受体"家族的两个原型成员。F1A α蛋白也会寡聚化。F1A α的过表达在哺乳动物细胞中诱导凋亡，并且Bcl-XL或FADD或半胱天冬酶-9的显性负突变体的共表达阻断了这种作用。缺失分析揭示了F1A α的中心区域，包括一组五个锚蛋白重复序列，这是最大凋亡活性所必需和足够的，N-末端区域似乎负调节这种活性。此外，F1A α在Asp（342）处被半胱天冬酶-3样蛋白酶切割，并且抗切割突变体在过表达时不能诱导细胞凋亡。因此，F1Aa是一个不断增长的死亡受体相关蛋白家族的成员，介导细胞凋亡。
Apoptosis is an evolutionarily conserved process that is critical for tissue homeostasis and development including sex determination in essentially all multicellular organisms. Here, we report the cloning of an ankyrin repeat-containing protein, termed F1A alpha, in a yeast two-hybrid screen using the cytoplasmic domain of Fas (CD95/APO-1) as bait. Amino acid sequence analysis indicates that F1A alpha has extensive homology to the sex-determining protein FEM-1 of the Caenorhabditis elegans, which is required for the development of all aspects of the male phenotype. F1A alpha associates with the cytoplasmic domains of Fas and tumor necrosis factor receptor 1, two prototype members of the "death receptor" family. The F1A alpha protein also oligomerizes. Overexpression of F1A alpha induces apoptosis in mammalian cells, and co-expression of Bcl-XL or the dominant negative mutants of either FADD or caspase-9 blocks this effect. Deletion analysis revealed the center region of F1A alpha, including a cluster of five ankyrin repeats to be necessary and sufficient for maximum apoptotic activity, and the N-terminal region appears to regulate negatively this activity. Furthermore, F1A alpha is cleaved by a caspase-3-like protease at Asp(342), and the cleavage-resistant mutant is unable to induce apoptosis upon overexpression. F1Aa is therefore a member of a growing family of death receptor-associated proteins that mediates apoptosis.