Phosphorylation of mitochondrial polyubiquitin by PINK1 promotes Parkin mitochondrial tethering.
Phosphorylation of mitochondrial polyubiquitin by PINK1 promotes Parkin mitochondrial tethering.
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DOI：
10.1371/journal.pgen.1004861
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发表时间：
2014-12
期刊：
影响因子：
4.5
通讯作者：
Imai Y
中科院分区：
文献类型：
--
作者：
Shiba-Fukushima K;Arano T;Matsumoto G;Inoshita T;Yoshida S;Ishihama Y;Ryu KY;Nukina N;Hattori N;Imai Y
PINK1和E3泛素(Ub)连接酶Parkin参与线粒体质量控制。PINK1使Parkin‘s泛素样区Ser65的磷酸化，刺激Parkin激活和移位到受损的线粒体，从而诱导有丝分裂生成PolyUb链。然而，Parkin Ser65的磷酸化对Parkin线粒体易位是不够的。在这里，我们报告了PolyUb链中的Ser65也被PINK1磷酸化，线粒体上磷酸化的PolyUb链束缚了线粒体上的Parkin。Tom70MTS-4xUb SE模拟线粒体上的磷酸化Ser65多聚Ub链的表达，激活了Parkin E3的活性及其线粒体易位。在Tom70MTS-4xUb SE存在下，E3-Dead形式的Parkin易位到线粒体，膜电位降低，而非磷酸多聚Ub突变体Tom70MTS-4xUb SA抑制了Parkin易位。Parkin通过其RING1环间(IBR)结构域与磷酸多聚Ub链结合，但其RING0连接子也是线粒体易位所必需的。此外，Tom70MTS-4xUb SE的表达改善了PINK1缺陷的果蝇的线粒体退化，但不能改善Parkin缺陷的果蝇的线粒体退化。我们的研究表明，PINK1对线粒体PolyUb的磷酸化参与了Parkin的激活和线粒体的易位，从而预测了线粒体磷酸化PolyUb的连锁反应机制，从而实现了Parkin的快速易位。帕金森氏病是一种神经退行性疾病，除了其他神经系统外，还会引起中脑多巴胺能系统的退化。PINK1和parkin分别编码线粒体蛋白激酶和胞浆Ub连接酶，是常染色体隐性遗传型青少年帕金森病的致病基因。线粒体膜电位降低时，PINK1的激活会从胞浆中招募Parkin，激活其Ub连接酶活性，从而确保通过有丝分裂去除受损的线粒体。然而，PINK1如何将帕金招募到受损的线粒体中仍不清楚。在这里，我们描述了PINK1对PolyUb链的磷酸化是在线粒体上招募Parkin的关键事件。Parkin与PINK1结合，并被Parkin与PINK1合作产生的磷酸多聚Ub激活。线粒体上磷酸化多聚Ub模拟蛋白的表达挽救了果蝇因PINK1缺失而导致的线粒体变性。我们的研究表明，存在Parkin激活和线粒体易位的放大级联反应，其中由PINK1和Parkin产生的线粒体上磷酸化的PolyUb的种子触发了Parkin募集和激活的连锁反应。
The kinase PINK1 and the E3 ubiquitin (Ub) ligase Parkin participate in mitochondrial quality control. The phosphorylation of Ser65 in Parkin's ubiquitin-like (UBl) domain by PINK1 stimulates Parkin activation and translocation to damaged mitochondria, which induces mitophagy generating polyUb chain. However, Parkin Ser65 phosphorylation is insufficient for Parkin mitochondrial translocation. Here we report that Ser65 in polyUb chain is also phosphorylated by PINK1, and that phosphorylated polyUb chain on mitochondria tethers Parkin at mitochondria. The expression of Tom70MTS-4xUb SE, which mimics phospho-Ser65 polyUb chains on the mitochondria, activated Parkin E3 activity and its mitochondrial translocation. An E3-dead form of Parkin translocated to mitochondria with reduced membrane potential in the presence of Tom70MTS-4xUb SE, whereas non-phospho-polyUb mutant Tom70MTS-4xUb SA abrogated Parkin translocation. Parkin binds to the phospho-polyUb chain through its RING1-In-Between-RING (IBR) domains, but its RING0-linker is also required for mitochondrial translocation. Moreover, the expression of Tom70MTS-4xUb SE improved mitochondrial degeneration in PINK1-deficient, but not Parkin-deficient, Drosophila. Our study suggests that the phosphorylation of mitochondrial polyUb by PINK1 is implicated in both Parkin activation and mitochondrial translocation, predicting a chain reaction mechanism of mitochondrial phospho-polyUb production by which rapid translocation of Parkin is achieved. Parkinson's disease is a neurodegenerative disorder caused by degeneration of the midbrain dopaminergic system in addition to other nervous systems. PINK1 and parkin, which encode mitochondrial protein kinase and cytosolic Ub ligase, respectively, were identified as the genes responsible for the autosomal recessive form of juvenile Parkinson's disease. Activation of PINK1 upon reduction of mitochondrial membrane potential recruits Parkin from the cytosol activating its Ub ligase activity, which ensures removal of damaged mitochondria through mitophagy. However, how PINK1 recruits Parkin to the damaged mitochondria remained unclear. Here, we describe that the phosphorylation of polyUb chain by PINK1 is a key event to recruit Parkin on the mitochondria. Parkin binds to, and is activated by, phospho-polyUb generated by Parkin in collaboration with PINK1. Expression of a phospho-polyUb mimetic protein on mitochondria rescued mitochondrial degeneration caused by loss of PINK1 in Drosophila. Our study suggests the existence of an amplification cascade of Parkin activation and mitochondrial translocation, in which a ‘seed' of phosphorylated polyUb on the mitochondria, generated by PINK1 and Parkin, triggers a chain reaction of Parkin recruitment and activation.
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影响因子：
4.5
作者：
Imai Y;Kanao T;Sawada T;Kobayashi Y;Moriwaki Y;Ishida Y;Takeda K;Ichijo H;Lu B;Takahashi R
通讯作者：
Takahashi R
DOI：
10.1073/pnas.0500346102
发表时间：
2005-07-19
影响因子：
11.1
作者：
Cha, GH;Kim, S;Cho, KS
通讯作者：
Cho, KS
DOI：
10.1083/jcb.201210111
发表时间：
2013-01-21
期刊：
The Journal of cell biology
影响因子：
--
作者：
Lazarou M;Narendra DP;Jin SM;Tekle E;Banerjee S;Youle RJ
通讯作者：
Youle RJ
影响因子：
4.8
作者：
Hristova, Ventzislava A.;Beasley, Steven A.;Shaw, Gary S.
通讯作者：
Shaw, Gary S.
DOI：
10.1083/jcb.200910140
发表时间：
2010-04-19
期刊：
The Journal of cell biology
影响因子：
--
作者：
Matsuda N;Sato S;Shiba K;Okatsu K;Saisho K;Gautier CA;Sou YS;Saiki S;Kawajiri S;Sato F;Kimura M;Komatsu M;Hattori N;Tanaka K
通讯作者：
Tanaka K