Hydrogen peroxide acts on sensitive mitochondrial proteins to induce death of a fungal pathogen revealed by proteomic analysis.
Hydrogen peroxide acts on sensitive mitochondrial proteins to induce death of a fungal pathogen revealed by proteomic analysis.
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蛋白质组学分析揭示过氧化氢作用于敏感线粒体蛋白以诱导真菌病原体死亡
DOI：
10.1371/journal.pone.0021945
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Tian S
中科院分区：
文献类型：
--
作者：
Qin G;Liu J;Cao B;Li B;Tian S
植物和动物的宿主细胞如何保护自己免受真菌入侵是一个生物学上有趣的和经济上重要的问题。在这里，我们调查的机械过程，导致死亡的扩展青霉，一种广泛的植物病原真菌，通过确定受过氧化氢（H2O2），这是经常产生的响应的宿主细胞的细胞化合物。我们表明，质膜损伤不是H2O2诱导的真菌病原体死亡的主要原因。蛋白质组学分析表明，在扩展青霉细胞总蛋白的变化中，线粒体可能参与了这一过程。然后，我们进行线粒体亚蛋白质组学分析，以寻找H2O2敏感的蛋白质，在扩展的P。鉴定了一组线粒体蛋白，包括呼吸链复合物I和III、F1F0 ATP合酶和线粒体磷酸盐载体蛋白。进一步研究了几种蛋白质的功能，以确定它们对H2O2诱导的真菌死亡的影响。通过荧光共定位和特异性抑制剂的使用，我们提供的证据表明，线粒体呼吸链的复合物III有助于ROS在真菌线粒体H2O2胁迫下的产生。ROS的不希望的积累引起线粒体蛋白的氧化损伤，并导致线粒体膜电位的崩溃。同时，我们证明ATP合酶参与真菌病原体对氧化胁迫的反应，因为寡霉素抑制ATP合酶会降低存活率。我们的数据表明，线粒体损伤由于氧化应激敏感蛋白的功能改变与真菌死亡引起的H2O2。
How the host cells of plants and animals protect themselves against fungal invasion is a biologically interesting and economically important problem. Here we investigate the mechanistic process that leads to death of Penicillium expansum, a widespread phytopathogenic fungus, by identifying the cellular compounds affected by hydrogen peroxide (H2O2) that is frequently produced as a response of the host cells. We show that plasma membrane damage was not the main reason for H2O2-induced death of the fungal pathogen. Proteomic analysis of the changes of total cellular proteins in P. expansum showed that a large proportion of the differentially expressed proteins appeared to be of mitochondrial origin, implying that mitochondria may be involved in this process. We then performed mitochondrial sub-proteomic analysis to seek the H2O2-sensitive proteins in P. expansum. A set of mitochondrial proteins were identified, including respiratory chain complexes I and III, F1F0 ATP synthase, and mitochondrial phosphate carrier protein. The functions of several proteins were further investigated to determine their effects on the H2O2-induced fungal death. Through fluorescent co-localization and the use of specific inhibitor, we provide evidence that complex III of the mitochondrial respiratory chain contributes to ROS generation in fungal mitochondria under H2O2 stress. The undesirable accumulation of ROS caused oxidative damage of mitochondrial proteins and led to the collapse of mitochondrial membrane potential. Meanwhile, we demonstrate that ATP synthase is involved in the response of fungal pathogen to oxidative stress, because inhibition of ATP synthase by oligomycin decreases survival. Our data suggest that mitochondrial impairment due to functional alteration of oxidative stress-sensitive proteins is associated with fungal death caused by H2O2.