Direct interaction between estrogen receptor α and NF-κB in the nucleus of living cells
Direct interaction between estrogen receptor α and NF-κB in the nucleus of living cells
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DOI：
10.1016/j.mce.2007.05.002
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发表时间：
2007-07-15
影响因子：
4.1
通讯作者：
Tertoolen, Leon G. J.
中科院分区：
文献类型：
--
作者：
Quaedackers, Monique E.;van den Brink, Christina E.;Tertoolen, Leon G. J.
类固醇受体对核因子-kappaB转录活性的抑制是类固醇激素抗炎作用的基础，这种相互作用的分子机制被认为涉及直接的蛋白质-蛋白质相互作用。在这项研究中，我们通过荧光共振能量转移(FRET)和免疫共沉淀，证明了雌激素受体(ER)α和核因子-kappaB在体内相互作用。用U2-OS细胞研究ERα荧光融合蛋白与核因子-kappaB亚单位p50和p65的直接相互作用。只有在细胞核中观察到相互作用，当ERα与NF-kappa B亚基共同表达时，FRET信号最大，细胞被雌激素刺激。这与在此条件下诱导蛋白质的核共定位是一致的。此外，在稳定表达的U2-OS克隆中，证实了与核因子-kappaB的相互作用。缺乏Rel同源结构域的p65缺失突变体在与ERα的相互作用中受到强烈破坏，表明该结构域的重要性。综上所述，这些发现为ERA和NF-kappa B之间的串扰的蛋白质-蛋白质直接相互作用模型提供了强有力的基础。(C)2007爱思唯尔爱尔兰有限公司。保留所有权利。
Inhibition of NF-kappa B transcriptional activity by steroid receptors is the basis for the antiinflammatory actions of steroid hormones and the molecular mechanism underlying this cross-talk is thought to involve direct protein-protein interactions. In this study, we show that estrogen receptor (ER)alpha and NF-kappa B interact in vivo by using fluorescence resonance energy transfer (FRET) and co-immunoprecipitation. U2-OS cells were used to study direct interactions between fluorescent fusion proteins of ER alpha and the NF-kappa B subunits p50 and p65. Interactions were observed only in the nucleus and maximal FRET signal was detected when ER alpha is co-expressed with both NF-kappa B subunits and cells were stimulated with estrogen. This is in agreement with the induction of nuclear co-localization of the proteins under this condition. Moreover, in a U2-OS clone stably expressing ERa, interaction with NF-kappa B was confirmed. A p65 deletion mutant lacking the Rel homology domain was strongly impaired in its interaction with ER alpha showing the importance of this domain. Taken together, these findings provide a strong basis for the direct protein-protein interaction model for cross-talk between ERa and NF-kappa B. (c) 2007 Elsevier Ireland Ltd. All rights reserved.