Targeted cell killing by reconstituted caspases
Targeted cell killing by reconstituted caspases
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DOI：
10.1073/pnas.0610877104
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发表时间：
2007-02-13
影响因子：
11.1
通讯作者：
Chalfie, Martin
中科院分区：
文献类型：
--
作者：
Chelur, Dattananda S.;Chalfie, Martin
我们开发了一种双组分系统，涉及重组半胱天冬酶（recCaspase），用于选择性和/或条件消融靶细胞。半胱天冬酶是程序性细胞死亡的刽子手，通常作为无活性的酶原合成，并通过其亚基的蛋白水解加工而激活。我们在这里展示了，使用两种不同的半胱天冬酶，秀丽隐杆线虫的CED-3和人类的caspase -3，亚基的共表达产生了组成性的半胱天冬酶活性，导致细胞死亡。然而，这种recCaspase活性仅在亚基通过连接的反平行亮氨酸拉链结构域结合时发生。我们利用recCaspases的双组分性质，通过表达启动子组合中的单个亚基，选择性地靶向细胞亚群进行凋亡，或在发育过程中的特定时间诱导特定细胞死亡。高度的靶特异性和对recCaspase诱导的严格调控将有利于创建用于特定细胞消融和其他靶向细胞杀伤的动物模型。
We have developed a two-component system involving reconstituted caspase (recCaspase) for selective and/or conditional ablation of targeted cells. Caspases, the executioners of programmed cell death, are normally synthesized as inactive zymogens and are activated by proteolytic processing of their subunits. We show here, using two different caspases, Caenorhabditis elegans CED-3 and human Caspase-3, that coexpression of the subunits generates constitutively active caspase activity that leads to cell death. This recCaspase activity, however, occurred only when the subunits associated through binding of linked antiparallel leucine-zipper domains. We exploited the dual-component nature of recCaspases by expressing the individual subunits from combinations of promoters either to target selectively the subset of cells for apoptosis or induce cell death in specific cells at specific times during development. The high degree of target specificity and tight regulation of induction of recCaspase would be advantageous in creating animal models that are ablated for specific cells and in other targeted cell killings.