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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >The NADPH-dependent thioredoxin system constitutes a functional backup for cytosolic glutathione reductase in Arabidopsis
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The NADPH-dependent thioredoxin system constitutes a functional backup for cytosolic glutathione reductase in Arabidopsis

机译：NADPH依赖的硫氧还蛋白系统构成拟南芥胞质谷胱甘肽还原酶的功能性后备

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摘要

Tight control of cellular redox homeostasis is essential for protection against oxidative damage and for maintenance of normal metabolism as well as redox signaling events. Under oxidative stress conditions, the tripeptide glutathione can switch from its reduced form (GSH) to oxidized glutathione disulfide (GSSG), and thus, forms an important cellular redox buffer. GSSG is normally reduced to GSH by 2 glutathione reductase (GR) isoforms encoded in the Arabidopsis genome, cytosolic GR1 and GR2 dual-targeted to chloroplasts and mitochondria. Measurements of total GR activity in leaf extracts of wild-type and 2 gr1 deletion mutants revealed that ≈65% of the total GR activity is attributed to GR1, whereas ≈35% is contributed by GR2. Despite the lack of a large share in total GR activity, gr1 mutants do not show any informative phe-notype, even under stress conditions, and thus, the physiological impact of GR1 remains obscure. To elucidate its role in plants, glutathione-specific redox-sensitive GFP was used to dynamically measure the glutathione redox potential (E_(GSH)) in the cytosol. Using this tool, it is shown that £gsh in gr1 mutants is significantly shifted toward more oxidizing conditions. Surprisingly, dynamic reduction of GSSG formed during induced oxidative stress in gr1 mutants is still possible, although significantly delayed compared with wild-type plants. We infer that there is functional redundancy in this critical pathway. Integrated biochemical and genetic assays identify the NADPH-dependent thioredoxin system as a backup system for GR1. Deletion of both, NADPH-dependent thioredoxin reductase A and GR1, prevents survival due to a pollen lethal phenotype.
机译：严格控制细胞氧化还原稳态对于防止氧化损伤，维持正常的代谢以及氧化还原信号传递事件至关重要。在氧化应激条件下，三肽谷胱甘肽可以从其还原形式（GSH）转变为氧化型谷胱甘肽二硫化物（GSSG），因此形成重要的细胞氧化还原缓冲液。 GSSG通常通过拟南芥基因组中编码的2种谷胱甘肽还原酶（GR）亚型，双重靶向叶绿体和线粒体的胞质GR1和GR2还原为GSH。对野生型和2个gr1缺失突变体叶片提取物中总GR活性的测量表明，总GR活性的约65％归功于GR1，而约35％的归因于GR2。尽管在总GR活性中缺乏很大份额，但gr1突变体即使在压力条件下也没有显示任何信息表型，因此，GR1的生理影响仍然不清楚。为了阐明其在植物中的作用，使用了谷胱甘肽特异性氧化还原敏感的GFP来动态测量细胞溶质中的谷胱甘肽氧化还原电位（E_（GSH））。使用该工具，表明gr1突变体中的£gsh显着向更多的氧化条件转移。出人意料的是，尽管与野生型植物相比明显延迟，但gr1突变体在诱导的氧化应激过程中形成的GSSG的动态降低仍然是可能的。我们推断此关键路径中存在功能冗余。综合的生化和遗传分析确定了依赖NADPH的硫氧还蛋白系统作为GR1的备用系统。删除NADPH依赖性硫氧还蛋白还原酶A和GR1均会阻止由于花粉致死表型而导致的存活。

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  • 作者

    Laurent Marty; Wafi Siala; Markus Schwarzlaender; Mark D. Fricker; Markus Wirtz; Lee J. Sweetlove; Yves Meyer; Andreas J. Meyer; Jean-Philippe Reichheld; Ruediger Hell;

  • 作者单位

    Heidelberg Institute for Plant Science, Heidelberg University, Im Neuenheimer Feld 360, 69120 Heidelberg, Germany;

    Laboratoire Genome et Developpement des Plantes, Unite Mixte de Recherche 5096, Centre National de la Recherche Scientifique/UP-lnstitut de Recherche pour le Developpement/Universite de Perpignan 52, Avenue Paul Alduy, 66860 Perpignan Cedex, France;

    Department of Plant Sciences, University of Oxford,South Parks Road, Oxford OX1 3RB, United Kingdom;

    Department of Plant Sciences, University of Oxford,South Parks Road, Oxford OX1 3RB, United Kingdom;

    Heidelberg Institute for Plant Science, Heidelberg University, Im Neuenheimer Feld 360, 69120 Heidelberg, Germany;

    Department of Plant Sciences, University of Oxford,South Parks Road, Oxford OX1 3RB, United Kingdom;

    Department of Plant Sciences, University of Oxford,South Parks Road, Oxford OX1 3RB, United Kingdom;

    Heidelberg Institute for Plant Science, Heidelberg University, Im Neuenheimer Feld 360, 69120 Heidelberg, Germany;

    Department of Plant Sciences, University of Oxford,South Parks Road, Oxford OX1 3RB, United Kingdom;

    Heidelberg Institute for Plant Science, Heidelberg University, Im Neuenheimer Feld 360, 69120 Heidelberg, Germany;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    redox homeostasis; redox imaging; redox-sensitive GFP; thioredoxin reductase;

    机译：氧化还原稳态氧化还原成像氧化还原敏感的GFP;硫氧还蛋白还原酶;

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