Beta tubulin affects the aryl hydrocarbon receptor function via an Arnt-mediated mechanism.
Beta tubulin affects the aryl hydrocarbon receptor function via an Arnt-mediated mechanism.
复制标题
β 微管蛋白通过 Arnt 介导的机制影响芳烃受体功能。
DOI：
10.1016/j.bcp.2009.12.010
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发表时间：
2010
影响因子：
5.8
通讯作者：
Chan,WilliamK
中科院分区：
文献类型：
--
作者：
Zhang,Tianmin;Wang,Xiaodong;Shinn,Annie;Jin,Jingjun;Chan,WilliamK
我们一直在研究在基因转录激活之前，芳烃受体核转位器（Arnt）依赖的DNA复合体形成的需要。采用DEAE色谱法，我们获得了富含β-微管蛋白的Sf9昆虫组分F5。F5抑制AhR凝胶移位复合物的形成，这种抑制对蛋白酶很敏感，这表明F5部分中存在的蛋白质是抑制的原因。另外的实验表明，在抗β-微管蛋白IgG存在时，这种抑制作用不那么明显，而猪脑中富含β-微管蛋白的部分也能抑制AhR凝胶移位的形成。Sf9 β-微管蛋白与Arnt相互作用，抑制AhR/Arnt异源二聚体与其相应增强子的结合。人β4-微管蛋白与Sf9 β-微管蛋白具有较高的序列同一性，通过降低细胞核中Arnt含量和保留细胞质中的Arnt来抑制ahr依赖性荧光素酶的表达。利用人β4-微管蛋白的GFP融合进行荧光研究发现，β4-微管蛋白可以阻止Arnt在Sf9细胞中的定位。我们提供的证据表明β-微管蛋白可能调节Arnt的生理含量。
We have been studying the requirement for the aryl hydrocarbon receptor nuclear translocator (Arnt)-dependent DNA complex formation, which precedes the activation of gene transcription. Using DEAE chromatography, we have obtained a Sf9 insect fraction F5 that is highly enriched with β-tubulin. F5 inhibits the formation of the AhR gel shift complex and this inhibition is sensitive to protease, suggesting that proteins that are present in this F5 fraction are responsible for the inhibition. Additional experiments have revealed that this inhibition is less pronounced in the presence of anti-β-tubulin IgG and β-tubulin enriched fraction from pig brain also inhibits the AhR gel shift formation. Sf9 β-tubulin interacts with Arnt and suppresses the binding of the AhR/Arnt heterodimer to its corresponding enhancer. Human β4-tubulin, which shares high sequence identity with Sf9 β-tubulin, suppresses the AhR-dependent luciferase expression by reducing the nuclear Arnt content and retaining Arnt in the cytoplasm. Fluorescence studies using the GFP fusion of human β4-tubulin have revealed that β4-tubulin prevents the localization of Arnt in Sf9 cells. Here we have provided evidence suggesting that β-tubulin may regulate the physiological content of Arnt.