Monomer DJ-1 and its N-terminal sequence are necessary for mitochondrial localization of DJ-1 mutants.
Monomer DJ-1 and its N-terminal sequence are necessary for mitochondrial localization of DJ-1 mutants.
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DOI：
10.1371/journal.pone.0054087
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Ariga H
中科院分区：
文献类型：
--
作者：
Maita C;Maita H;Iguchi-Ariga SM;Ariga H
DJ-1是一种新的癌基因，也是家族性帕金森病（park 7）的致病基因。DJ-1具有转录调节、抗氧化反应、分子伴侣和线粒体调节等多种功能。在DJ-1基因敲除小鼠和鱼苗中观察到线粒体功能障碍，并且线粒体DJ-1对氧化应激诱导的细胞死亡更具保护作用。虽然DJ-1易位到线粒体中被氧化应激增强，导致DJ-1的半胱氨酸106（C106）氧化，但线粒体DJ-1的特性和DJ-1易位到线粒体中的机制知之甚少。在这项研究中，免疫染色，免疫共沉淀，细胞分级分离和下拉实验表明，谷氨酰胺18（E18）DJ-1的突变体定位于线粒体，不使同源二聚体。同样，发现具有位于二聚体界面的两个半胱氨酸C46 S和C53 A突变的DJ-1以及致病突变体M26 I和L166 P DJ-1定位于线粒体中并且不产生同源二聚体。含有E18 A和C106 S（其中C106未被氧化）的突变体DJ-1也定位在线粒体中，这表明C106的氧化对于DJ-1的线粒体定位很重要，但不是必需的。应该注意的是，当通过用CCCP（线粒体中氧化磷酸化系统的解偶联剂）处理细胞而降低线粒体膜电位时，E18 A DJ-1从线粒体易位至细胞质。此外，N端12个氨基酸的缺失或取代导致E18 A、M26 I和L166 P DJ-1从线粒体重新定位到细胞质中。这些结果表明，单体和N-末端12个氨基酸是必需的DJ-1突变体的线粒体定位和构象变化引起的C106氧化或E18突变导致的DJ-1易位到线粒体。
DJ-1 is a novel oncogene and also a causative gene for familial Parkinson’s disease (park7). DJ-1 has multiple functions that include transcriptional regulation, anti-oxidative reaction and chaperone and mitochondrial regulation. Mitochondrial dysfunction is observed in DJ-1-knockout mice and fry, and mitochondrial DJ-1 is more protective against oxidative stress-induced cell death. Although translocation of DJ-1 into mitochondria is enhanced by oxidative stress that leads to oxidation of cysteine 106 (C106) of DJ-1, the characteristics of mitochondrial DJ-1 and the mechanism by which DJ-1 is translocated into mitochondria are poorly understood. In this study, immunostaining, co-immunoprecipitation, cell fractionation and pull-down experiments showed that mutants of glutamine 18 (E18) DJ-1 are localized in mitochondria and do not make homodimers. Likewise, DJ-1 with mutations of two cysteines located in the dimer interface, C46S and C53A, and pathogenic mutants, M26I and L166P DJ-1, were found to be localized in mitochondria and not to make homodimers. Mutant DJ-1 harboring both E18A and C106S, in which C106 is not oxidized, was also localized in mitochondria, indicating that oxidation of C106 is important but not essential for mitochondrial localization of DJ-1. It should be noted that E18A DJ-1 was translocated from mitochondria to the cytoplasm when mitochondrial membrane potential was reduced by treatment of cells with CCCP, an uncoupler of the oxidative phosphorylation system in mitochondria. Furthermore, deletion or substitution of the N-terminal 12 amino acids in DJ-1 resulted in re-localization of E18A, M26I and L166P DJ-1 from mitochondria into the cytoplasm. These findings suggest that a monomer and the N-terminal 12 amino acids are necessary for mitochondrial localization of DJ-1 mutants and that conformation change induced by C106 oxidation or by E18 mutation leads to translocation of DJ-1 into mitochondria.
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DOI：
10.1073/pnas.0911175107
发表时间：
2010-05-25
影响因子：
11.1
作者：
Hao, Ling-Yang;Giasson, Benoit I.;Bonini, Nancy M.
通讯作者：
Bonini, Nancy M.
影响因子：
4.8
作者：
Honbou, K;Suzuki, NN;Inagaki, F
通讯作者：
Inagaki, F
影响因子：
56.9
作者：
Bonifati, V;Rizzu, P;Heutink, P
通讯作者：
Heutink, P
DOI：
10.1073/pnas.0607260103
发表时间：
2006-10-10
影响因子：
11.1
作者：
Clements, Casey M.;McNally, Richard S.;Ting, Jenny P-Y.
通讯作者：
Ting, Jenny P-Y.
影响因子：
2.9
作者：
Hulleman, John D.;Mirzaei, Hamid;Rochet, Jean-Christophe
通讯作者：
Rochet, Jean-Christophe