Identification of the interactive interface and phylogenic conservation of the Nrf2-Keap1 system
Identification of the interactive interface and phylogenic conservation of the Nrf2-Keap1 system
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DOI：
10.1046/j.1365-2443.2002.00561.x
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发表时间：
2002-08-01
期刊：
影响因子：
2.1
通讯作者：
Yamamoto, M
中科院分区：
文献类型：
--
作者：
Kobayashi, M;Itoh, K;Yamamoto, M
背景：转录因子Nrf2及其负调控因子Keap1在一系列解毒和抗氧化酶的转录诱导中发挥重要作用。为了深入了解细胞如何接收氧化和亲电信号并将其转导到Nrf2，我们开发了一个斑马鱼模型系统用于分子毒理学研究。结果：系统克隆了斑马鱼细胞保护酶cdna，发现亲电剂能有效诱导其表达。因此，我们确定了Nrf2和Keap1在斑马鱼中的存在。功能缺失和功能获得分析表明，Nrf2是细胞保护酶基因亚群的主要调节因子，而Keap1在斑马鱼中抑制Nrf2的活性。通过反向双杂交筛选，在Nrf2的Neh2结构域中发现了一个对Nrf2- keap1相互作用至关重要的ETGE基序，并发现该基序对于调控Nrf2在斑马鱼中的活性是必不可少的。结论：综上所述，Nrf2-Keap1系统在脊椎动物中具有高度保守性，Nrf2与Keap1之间的界面构成了该调控系统的重要分子基础。
Background: The transcription factor Nrf2 and its negative regulator Keap1 play important roles in transcriptional induction of a set of detoxifying and anti-oxidant enzymes. To gain an insight into our present enigma as to how cells receive oxidative and electrophilic signals and transduce them to Nrf2, we have developed a zebrafish model system for molecular toxicological studies.Results: We systematically cloned zebrafish cytoprotective enzyme cDNAs and found their expression to be efficiently induced by electrophilic agents. We consequently identified the presence of Nrf2 and Keap1 in zebrafish. Both loss- and gain-of-function analyses demonstrated that Nrf2 is the primary regulator of a subset of cytoprotective enzyme genes, while Keap1 suppresses Nrf2 activity in zebrafish. An ETGE motif, critical for the Nrf2-Keap1 interaction, was identified in the Neh2 domain of Nrf2 by reverse two-hybrid screening and found to be indispensable for the regulation of Nrf2 activity in zebrafish.Conclusion: Taken together, these results indicate that the Nrf2-Keap1 system is highly conserved among vertebrates and that the interface between Nrf2 and Keap1 forms an important molecular basis of this regulatory system.