C. elegans MAC-1, an essential member of the AAA family of ATPases, can bind CED-4 and prevent cell death.
C. elegans MAC-1, an essential member of the AAA family of ATPases, can bind CED-4 and prevent cell death.
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发表时间：
1999-05
期刊：
影响因子：
4.6
通讯作者：
D. Wu;P. Chen;S. Chen;Y. Hu;G. Núñez;R. Ellis
中科院分区：
文献类型：
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作者：
D. Wu;P. Chen;S. Chen;Y. Hu;G. Núñez;R. Ellis
在秀丽隐杆线虫中，CED-4在程序性细胞死亡的调控中起核心作用。为了鉴定具有必要或多效性活性的蛋白，这些蛋白也可能调节细胞死亡，我们使用酵母双杂交系统筛选ced -4结合蛋白。我们确定了MAC-1，它是atp酶AAA家族的一员，类似于果蝇的Smallminded。免疫沉淀研究证实，MAC-1与CED-4相互作用，也与CED-4的哺乳动物同源物Apaf-1相互作用。此外，MAC-1可以形成包括CED-3或CED-9在内的多蛋白复合物。在热休克启动子控制下的MAC-1转基因可防止秀丽隐杆线虫的一些自然细胞死亡，并且这种保护在ced-9(n1950sd)/+遗传背景下得到增强。我们在哺乳动物细胞中观察到类似的效果，在哺乳动物细胞中，MAC-1的表达可以阻止CED-4和CED-3诱导细胞凋亡。最后，mac-1是一个必不可少的基因，因为rna介导的干扰使蠕虫在幼虫发育早期失活。这种阻滞与在Smallminded突变体中观察到的相似，但与MAC-1结合CED-4的能力无关，因为它仍然发生在ced-3或CED-4零突变体中。这些结果表明，MAC-1识别了一类新的蛋白质，这些蛋白质对发育至关重要，并且可能在特定情况下调节细胞死亡。
In the nematode Caenorhabditis elegans, CED-4 plays a central role in the regulation of programmed cell death. To identify proteins with essential or pleiotropic activities that might also regulate cell death, we used the yeast two-hybrid system to screen for CED-4-binding proteins. We identified MAC-1, a member of the AAA family of ATPases that is similar to Smallminded of Drosophila. Immunoprecipitation studies confirm that MAC-1 interacts with CED-4, and also with Apaf-1, the mammalian homologue of CED-4. Furthermore, MAC-1 can form a multi-protein complex that also includes CED-3 or CED-9. A MAC-1 transgene under the control of a heat shock promoter prevents some natural cell deaths in C. elegans, and this protection is enhanced in a ced-9(n1950sd)/+ genetic background. We observe a similar effect in mammalian cells, where expression of MAC-1 can prevent CED-4 and CED-3 from inducing apoptosis. Finally, mac-1 is an essential gene, since inactivation by RNA-mediated interference causes worms to arrest early in larval development. This arrest is similar to that observed in Smallminded mutants, but is not related to the ability of MAC-1 to bind CED-4, since it still occurs in ced-3 or ced-4 null mutants. These results suggest that MAC-1 identifies a new class of proteins that are essential for development, and which might regulate cell death in specific circumstances.