Gene expression profiling reveals a signaling role of glutathione in redox regulation
Gene expression profiling reveals a signaling role of glutathione in redox regulation
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DOI：
10.1073/pnas.0504398102
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发表时间：
2005-09-27
影响因子：
11.1
通讯作者：
Ghezzi, P
中科院分区：
文献类型：
--
作者：
Fratelli, M;Goodwin, LO;Ghezzi, P
蛋白质可以与谷胱甘肽（GSH）形成可逆的混合二硫键。据推测，蛋白质谷胱甘肽化可能代表氧化还原调节的机制，以类似于蛋白质磷酸化介导的方式。我们研究了GSH除了其明显的抗氧化作用外，是否在HL 60细胞对过氧化氢（11202）的反应中具有信号传导作用。我们确定了早期的基因表达的变化，在不同的时间诱导过氧化氢处理，在增加蛋白谷胱甘肽化和最小的毒性条件下。然后，我们研究了先前的GSH消耗丁硫氨酸亚砜和马来酸二乙酯对这种反应的影响。分析显示2,016个基因受H2 O2调节，其中215个基因表现出GSH依赖性表达变化，可分为4个簇，表现出H2 O2的下调或上调，或被GSH耗尽增强或抑制。20个基因的调控通过实时RT-PCR进行验证。在最大的簇中过度表达的生物学过程类别（其上调被GSH耗竭抑制的基因）是NF-κ B B激活、转录和DNA甲基化。该簇还包括几种细胞因子和趋化因子配体和受体、氧化还原调节剂硫氧还蛋白相互作用蛋白和组蛋白去乙酰化酶sirtuin。其上调由GSH耗竭增强的基因簇包括两个HSP（HISP 40和HISP 70）和AP-1转录因子组分Fos和FosB。这项工作表明，谷胱甘肽，除了其抗氧化剂和保护功能，对氧化应激，有一个特定的信号作用，在氧化还原调节。
Proteins can form reversible mixed disulficles with glutathione (GSH). It has been hypothesized that protein glutathionylation may represent a mechanism of redox regulation, in a fashion similar to that mediated by protein phosphorylation. We investigated whether GSH has a signaling role in the response of HL60 cells to hydrogen peroxide (11202), in addition to its obvious antioxiclant role. We identified early changes in gene expression induced at different times by H2O2 treatment, under conditions that increase protein glutathionylation and minimal toxicity. We then investigated the effect of prior GSH depletion by buthionine sulfoximine and diethylmaleate on this response. The analysis revealed 2,016 genes regulated by H2O2- Of these, 215 genes showed GSH-dependent expression changes, classifiable into four clusters displaying down- or up-regulation by H2O2, either potentiated or inhibited by GSH depletion. The modulation of 20 selected genes was validated by real-time RT-PCR. The biological process categories overrepresented in the largest cluster (genes whose up-regulation was inhibited by GSH depletion) were NF-kappa B activation, transcription, and DNA methylation. This cluster also included several cytokine and chemokine ligands and receptors, the redox regulator thioredoxin interacting protein, and the histone deacetylase sirtuin. The cluster of genes whose up-regulation was potentiated by GSH depletion included two HSPs (HISP40 and HISP70) and the AP-1 transcription factor components Fos and FosB. This work demonstrates that GSH, in addition to its antioxiclant and protective function against oxidative stress, has a specific signaling role in redox regulation.