Nek1 regulates cell death and mitochondrial membrane permeability through phosphorylation of VDAC1.
Nek1 regulates cell death and mitochondrial membrane permeability through phosphorylation of VDAC1.
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DOI：
10.4161/cc.8.2.7551
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发表时间：
2009-01-15
期刊：
影响因子：
--
通讯作者：
Riley DJ
中科院分区：
文献类型：
--
作者：
Chen Y;Craigen WJ;Riley DJ
哺乳动物NIMA相关蛋白激酶1（Nek 1）对于DNA损伤后保持细胞存活非常重要，但损伤细胞在没有功能性Nek 1的情况下死亡的机制尚未得到证实。在这里，我们表明，Nek 1调节线粒体细胞死亡的途径，通过磷酸化的电压依赖性阴离子通道1（VDAC 1）的丝氨酸193。Nek 1与VDAC 1在酵母双杂交系统中，以及通过GST下拉测定和通过相互免疫沉淀。细胞中Nek 1的一部分也定位于线粒体。激酶死亡的Nek 1突变体的异位表达导致细胞死亡，其之前立即发生Nek 1依赖性VDAC 1-S193磷酸化的丧失。Nek 1缺陷细胞的UV照射或内源性Nek 1表达的沉默类似地导致细胞死亡前特异性S193磷酸化的丧失。Nek 1缺陷细胞的特征在于过度的线粒体膜通透性（MMP）和加速的细胞死亡。不能被Nek 1磷酸化的VDAC 1-Ser 193 Ala突变体的异位表达也导致细胞死亡。一个VDAC 1-Ser 193 Glu突变体，旨在模拟由Nek 1组成的磷酸化，拯救夸大的MMP，并保持细胞在DNA损伤损伤后存活，但只是短暂的。Nek 1和VDAC 1之间的直接相互作用提供了一种解释Nek 1如何防止过度细胞死亡的机制，以及特定激酶调节VDAC 1活性的第一个直接证据。
The mammalian NIMA-related protein kinase 1 (Nek1) is important for keeping cells alive after DNA damage, but the mechanism by which injured cells die without functional Nek1 has not yet been demonstrated. Here we show that Nek1 regulates the pathway to mitochondrial cell death through phosphorylation of voltage dependent anion channel 1 (VDAC1) on serine 193. Nek1 associates with VDAC1 in a yeast two-hybrid system, as well as by GST pull-down assays and by reciprocal immunoprecipitation. A portion of Nek1 in cells also localizes at mitochondria. Ectopic expression of a kinase-dead Nek1 mutant results in cell death, which is immediately preceded by loss of the Nek1-dependent VDAC1-S193 phosphorylation. UV irradiation of Nek1-deficient cells or silencing of endogenous Nek1 expression similarly results in loss of the specific S193 phosphorylation before cells die. Nek1-deficient cells are characterized by exaggerated mitochondrial membrane permeability (MMP) and accelerated cell death. Ectopic expression of a VDAC1-Ser193Ala mutant, which cannot be phosphorylated by Nek1, also results in cell death. A VDAC1-Ser193Glu mutant, designed to mimic constitutive phosphorylation by Nek1, rescues exaggerated MMP and keeps cells alive after DNA damaging injury, but only transiently. The direct interaction between Nek1 and VDAC1 provides a mechanism to explain how Nek1 prevents excessive cell death, as well as the first direct evidence that a specific kinase regulates VDAC1 activity.
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DOI：
10.1016/j.bbamcr.2006.01.009
发表时间：
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影响因子：
5.1
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