A stress-responsive glutathione S-transferase confers resistance to oxidative stress in Caenorhabditis elegans
A stress-responsive glutathione S-transferase confers resistance to oxidative stress in Caenorhabditis elegans
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DOI：
10.1016/s0891-5849(03)00102-3
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发表时间：
2003-06-01
影响因子：
7.4
通讯作者：
Henkle-Dührsen, K
中科院分区：
文献类型：
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作者：
Leiers, BR;Kampkötter, A;Henkle-Dührsen, K
先前的研究表明，秀丽隐杆线虫 GST-p24 在氧化应激反应中在稳态 mRNA 水平上调 [1]。本研究通过分析转基因线虫菌株 CL2166 和 CL3166 中的 Ce-GST-p24 启动子-报告基因结构，证实了转录上调。转基因菌株 BL1 过表达 Ce-GST-p24 酶（作为由其自身启动子控制的 GFP 融合蛋白），旨在研究该酶的体内功能。与野生型相比，BL1 的应激实验表明，其对细胞内诱导的氧化应激的抵抗力有所增强。通过对 BL1 线虫进行 RNAi 处理以沉默内基因和转基因 Ce-GST-p24，并通过分析 K08F4.7 纯合缺失突变体来检查 Ce-GST-p24 酶浓度降低的后果。在这两种情况下，Ce-GST-p24酶水平的降低都会导致线虫的抗逆性显着下降。这些结果清楚地证明了 Ce-GST-p24 浓度与氧化应激抵抗力之间的直接相关性。我们首次证明，操纵单个 GST 的表达可以调节机体对氧化应激的反应。用各种底物检查了这种解毒酶的酶活性，重点关注脂质过氧化产物。通过共焦激光扫描显微镜，Ce-GST-p24 也定位于 BL1 线虫中，揭示了广泛的分布特征。 (C) 2003 爱思唯尔公司。
Previous studies demonstrated that the Caenorhabditis elegans GST-p24 is upregulated at the steady state mRNA level in response to oxidative stress [1]. A transcriptional upregulation was confirmed in the current study by analyzing Ce-GST-p24 promoter-reporter constructs in transgenic C elegans strains CL2166 and CL3166. The transgenic strain BL1, which overexpresses the Ce-GST-p24 enzyme (as a GFP fusion protein controlled by its own promoter), was generated to investigate the function of this enzyme in vivo. Stress experiments with BL1 demonstrated an increased resistance to intracellularly induced oxidative stress, as compared to wild type. The consequences of a decrease in the Ce-GST-p24 enzyme concentration were examined by RNAi-treatment of BL1 C. elegans to silence both the endogene and the transgene Ce-GST-p24 and by the analysis of the K08F4.7 homozygous deletion mutant. In both cases, the reduced Ce-GST-p24 enzyme level resulted in a significant decrease in the stress resistance of the nematodes. These results clearly demonstrate a direct correlation between the concentration of Ce-GST-p24 and the resistance to oxidative stress. We have demonstrated for the first time that manipulation of the expression of a single GST can modulate the organismal reponse to oxidative stress. The enzymatic activity of this detoxification enzyme was examined with various substrates, giving emphasis to lipid peroxidation products. The Ce-GST-p24 was also localized in BL1 C. elegans by confocal laser-scanning microscopy, revealing a wide-spread distribution profile. (C) 2003 Elsevier Inc.