Tetraspanin is required for generation of reactive oxygen species by the dual oxidase system in Caenorhabditis elegans.
Tetraspanin is required for generation of reactive oxygen species by the dual oxidase system in Caenorhabditis elegans.
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DOI：
10.1371/journal.pgen.1002957
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发表时间：
2012-09
期刊：
影响因子：
4.5
通讯作者：
Mekada E
中科院分区：
文献类型：
--
作者：
Moribe H;Konakawa R;Koga D;Ushiki T;Nakamura K;Mekada E
活性氧（ROS）是一种有毒的分子，是机体防御和细胞信号转导的重要分子.保守的NADPH氧化酶（NOX）家族酶指导ROS的调节产生。过氧化氢（H2 O2）产生的双重氧化酶（DUOXs），一个成员的NOX家族，是至关重要的先天性粘膜免疫。此外，H2 O2是蛋白质修饰介导的细胞信号传导所必需的，例如哺乳动物中的甲状腺激素生物合成途径。与其他NOX同工酶相比，DUOX活性的调节机制尚不清楚。使用秀丽隐杆线虫作为模型，我们表明，四跨膜蛋白是所需的DUOX信号通路的诱导与双氧化酶成熟因子（DUOXA）。在当前的研究中，我们发现C. DUOX（bli-3）、DUOXA（doxa-1）和过氧化物酶（mlt-7）的基因突变。秀丽线虫引起与四跨膜蛋白TSP-15突变体相同的缺陷，表现为由于胶原蛋白的酪氨酸交联失败而引起的外骨骼缺陷。bli-3和doxa-1的共表达恢复了tsp-15突变体中的缺陷，表明tsp-15参与了ROS的产生。当在哺乳动物细胞中重组时，BLI-3产生H2 O2完全依赖于TSP-15。我们还证明了TSP-15、BLI-3和DOXA-1在体外和体内形成复合物。基于细胞融合的分析表明，细胞表面与TSP-15的结合对于BLI-3激活释放H2 O2至关重要。这项研究为四跨膜蛋白在ROS生成中的重要作用提供了第一个证据。ROS是高度反应性的分子，其可以在有氧代谢期间或通过外源性应激（例如暴露于UV光和辐射）不适当地产生。ROS与包括核酸、脂质和蛋白质在内的细胞组分相互作用，并不可逆地抑制其功能。然而，活性氧是先天宿主防御和多种生理过程所必需的，并由保守的NADPH氧化酶（NOX）家族酶产生。必须适当控制ROS生成酶释放ROS，因为慢性氧化应激可导致氧化还原状态失衡，通常与疾病和衰老有关。利用C.以线虫为模型，我们鉴定了四跨膜蛋白（TSP-15）作为双氧化酶（BLI-3）控制的ROS生成系统的一个新的关键组分，双氧化酶（BLI-3）是线虫中独特的NOX同工酶。优美的bli-3和tsp-15的突变体在细胞外基质交联中产生了相同的缺陷。在哺乳动物细胞中使用遗传学和重建实验的组合，我们已经证明了一种新的要求四跨膜蛋白的双氧化酶依赖性的活性氧生成通过复杂的形成在细胞表面。
Reactive oxygen species (ROS) are toxic but essential molecules responsible for host defense and cellular signaling. Conserved NADPH oxidase (NOX) family enzymes direct the regulated production of ROS. Hydrogen peroxide (H2O2) generated by dual oxidases (DUOXs), a member of the NOX family, is crucial for innate mucosal immunity. In addition, H2O2 is required for cellular signaling mediated by protein modifications, such as the thyroid hormone biosynthetic pathway in mammals. In contrast to other NOX isozymes, the regulatory mechanisms of DUOX activity are less understood. Using Caenorhabditis elegans as a model, we demonstrate that the tetraspanin protein is required for induction of the DUOX signaling pathway in conjunction with the dual oxidase maturation factor (DUOXA). In the current study, we show that genetic mutation of DUOX (bli-3), DUOXA (doxa-1), and peroxidase (mlt-7) in C. elegans causes the same defects as a tetraspanin tsp-15 mutant, represented by exoskeletal deficiencies due to the failure of tyrosine cross-linking of collagen. The deficiency in the tsp-15 mutant was restored by co-expression of bli-3 and doxa-1, indicating the involvement of tsp-15 in the generation of ROS. H2O2 generation by BLI-3 was completely dependent on TSP-15 when reconstituted in mammalian cells. We also demonstrated that TSP-15, BLI-3, and DOXA-1 form complexes in vitro and in vivo. Cell-fusion-based analysis suggested that association with TSP-15 at the cell surface is crucial for BLI-3 activation to release H2O2. This study provides the first evidence for an essential role of tetraspanin in ROS generation. ROS are highly reactive molecules, which can be inappropriately produced during aerobic metabolism or by exogenous stresses such as exposure to UV light and radiation. ROS interact with cellular components including nucleic acids, lipids, and proteins and irreversibly inhibit their functions. However, ROS are essential for innate host defense and multiple physiological processes and are generated by conserved NADPH oxidase (NOX) family enzymes. The release of ROS by ROS generator enzymes must be properly controlled, as chronic oxidative stress can cause an imbalance of the redox state and is often associated with disease and aging. Using C. elegans as a model, we identified a tetraspanin (TSP-15) protein as a new key component of the ROS generation system controlled by dual oxidase (BLI-3), a unique NOX isozyme in C. elegans. Mutants of both bli-3 and tsp-15 developed the same defects in extracellular matrix cross-linking. Using a combination of genetics and reconstitution experiments in mammalian cells, we have demonstrated a novel requirement of tetraspanin for dual oxidase-dependent ROS generation via complex formation at the cell surface.
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影响因子：
4.8
作者：
Ameziane-El-Hassani, R;Morand, S;Dupuy, C
通讯作者：
Dupuy, C
影响因子：
4.8
作者：
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影响因子：
3.7
作者：
De Deken, X;Wang, DT;Miot, F
通讯作者：
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DOI：
10.1083/jcb.201102095
发表时间：
2011-07-11
期刊：
The Journal of cell biology
影响因子：
--
作者：
Finkel T
通讯作者：
Finkel T
影响因子：
4.5
作者：
Benedetto, Alexandre;Au, Catherine;Aschner, Michael
通讯作者：
Aschner, Michael