PINK1-mediated phosphorylation of Parkin boosts Parkin activity in Drosophila.
PINK1-mediated phosphorylation of Parkin boosts Parkin activity in Drosophila.
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DOI：
10.1371/journal.pgen.1004391
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发表时间：
2014-06
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;Inoshita T;Hattori N;Imai Y
与早发性帕金森氏症相关的两个基因PINK1和Parkin分别编码一种蛋白激酶和一种泛素连接酶。这两种酶都被认为支持线粒体质量控制。我们已经报道，在哺乳动物培养细胞中，Parkin被PINK1磷酸化，位于泛素样域的Ser65位。然而，目前尚不清楚Parkin磷酸化是否参与体内线粒体的维持和多巴胺能神经元的活动。在这里，我们研究了Parkin磷酸化在果蝇中的作用，其中磷酸化残基在Ser94处保守。野生型Parkin异位表达引起的肌肉组织和脑多巴胺能神经元线粒体的形态变化在PINK1缺失时消失。相反，无论PINK1活性如何，拟磷Parkin都加速了线粒体的碎裂或聚集以及线粒体蛋白质的降解，这表明Parkin的磷酸化增强了其泛素连接酶的活性。一种非磷酸化形式的Parkin完全挽救了由于PINK1活性丧失而导致的肌肉线粒体退化，而在Parkin缺失果蝇中引入非磷酸化Parkin突变体导致肌肉组织中出现异常融合的线粒体。控制Parkin的磷酸化状态影响多巴胺能神经元神经末梢的自发多巴胺释放、多巴胺能神经元的存活率和飞行活动。我们的数据显示，Parkin磷酸化不仅调节线粒体功能，而且调节体内多巴胺能神经元的活动，提示适当的Parkin磷酸化调节对肌肉和多巴胺能功能是重要的。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分别编码蛋白激酶和泛素连接酶，被鉴定为常染色体隐性遗传型青少年帕金森病的致病基因。这两种酶与线粒体的维持有关。虽然我们之前在哺乳动物培养细胞中发现PINK1使Parkin磷酸化，但这种相互作用在体内的生理意义尚不清楚。在这里，我们描述了Parkin的磷酸化改变了肌肉组织中线粒体的形态和功能，这是通过降解线粒体GTP酶蛋白(如Mitofusin和Miro)和线粒体呼吸复合体I亚基，增加其泛素连接酶的活性来实现的。我们还发现，成分磷酸化和非磷酸化形式的Parkin的多巴胺能表达影响苍蝇的飞行活动并缩短寿命，这表明适当的Parkin磷酸化对多巴胺能活动和多巴胺能神经元的生存都是重要的。
Two genes linked to early onset Parkinson's disease, PINK1 and Parkin, encode a protein kinase and a ubiquitin-ligase, respectively. Both enzymes have been suggested to support mitochondrial quality control. We have reported that Parkin is phosphorylated at Ser65 within the ubiquitin-like domain by PINK1 in mammalian cultured cells. However, it remains unclear whether Parkin phosphorylation is involved in mitochondrial maintenance and activity of dopaminergic neurons in vivo. Here, we examined the effects of Parkin phosphorylation in Drosophila, in which the phosphorylation residue is conserved at Ser94. Morphological changes of mitochondria caused by the ectopic expression of wild-type Parkin in muscle tissue and brain dopaminergic neurons disappeared in the absence of PINK1. In contrast, phosphomimetic Parkin accelerated mitochondrial fragmentation or aggregation and the degradation of mitochondrial proteins regardless of PINK1 activity, suggesting that the phosphorylation of Parkin boosts its ubiquitin-ligase activity. A non-phosphorylated form of Parkin fully rescued the muscular mitochondrial degeneration due to the loss of PINK1 activity, whereas the introduction of the non-phosphorylated Parkin mutant in Parkin-null flies led to the emergence of abnormally fused mitochondria in the muscle tissue. Manipulating the Parkin phosphorylation status affected spontaneous dopamine release in the nerve terminals of dopaminergic neurons, the survivability of dopaminergic neurons and flight activity. Our data reveal that Parkin phosphorylation regulates not only mitochondrial function but also the neuronal activity of dopaminergic neurons in vivo, suggesting that the appropriate regulation of Parkin phosphorylation is important for muscular and dopaminergic functions. 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 protein kinase and ubiquitin-ligase, respectively, were identified as the genes responsible for the autosomal recessive form of juvenile Parkinson's disease. These two enzymes are involved in mitochondrial maintenance. Although we previously found that Parkin is phosphorylated by PINK1 in mammalian cultured cells, the physiological significance of this interaction in vivo remained unclear. Here, we describe that the phosphorylation of Parkin altered mitochondrial morphology and function in muscle tissue through the degradation of mitochondrial GTPase proteins (such as Mitofusin and Miro) and a mitochondrial respiratory complex I subunit by increasing its ubiquitin-ligase activity. We also found that the dopaminergic expression of both constitutively phosphorylated and non-phosphorylated forms of Parkin affects the flight activity and shortens the lifespan of flies, suggesting that the appropriate phosphorylation of Parkin is important for both dopaminergic activity and the survival of dopaminergic neurons.
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影响因子：
4.5
作者：
Liu S;Sawada T;Lee S;Yu W;Silverio G;Alapatt P;Millan I;Shen A;Saxton W;Kanao T;Takahashi R;Hattori N;Imai Y;Lu B
通讯作者：
Lu B
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：
10.1083/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
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
影响因子：
21.3
作者：
Fallon, Lara;Belanger, Catherine M. L.;Fon, Edward A.
通讯作者：
Fon, Edward A.