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首页> 美国卫生研究院文献>Biochemical Journal >Role of hepatocyte nuclear factor-3 alpha and hepatocyte nuclear factor-3 beta in Clara cell secretory protein gene expression in the bronchiolar epithelium.
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Role of hepatocyte nuclear factor-3 alpha and hepatocyte nuclear factor-3 beta in Clara cell secretory protein gene expression in the bronchiolar epithelium.

机译：肝细胞核因子3α和肝细胞核因子3β在细支气管上皮细胞Clara细胞分泌蛋白基因表达中的作用。

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摘要

The 5' flanking region of the Clara cell secretory protein (CCSP) gene contains two cis-acting elements which bind hepatocyte nuclear factor (HNF)-3 alpha and HNF-3 beta in vitro. To determine the role of these proteins in mediating CCSP gene expression in the bronchiolar epithelium, chimeric CCSP-reporter gene constructs containing various regions of the CCSP 5' flanking region were co-transfected into H-441 cells with HNF-3 alpha or HNF-3 beta expression plasmids. These studies indicate that each of these transcription factors positively regulates CCSP gene expression and revealed that CCSP region I (-132 to -76) is sufficient to mediate this effect. Gel-mobility-shift assays with oligonucleotides corresponding to CCSP region I, nuclear extract from bronchiolar epithelial cells and HNF-3-specific antibodies indicate that HNF-3 alpha and HNF-3 beta are the only proteins in bronchiolar epithelial cells which directly interact with this region. Consistent with these observations, HNF-3 alpha and HNF-3 beta transcripts were found to be enriched in this cell population and in situ hybridization of adult lung revealed HNF-3 gene expression in non-ciliated bronchiolar epithelial cells expressing the CCSP gene. Finally, experiments with CCSP region I and a heterologous promoter indicate that this region acts in a promoter-specific context, suggesting that additional factors interacting via the minimal CCSP promoter region are essential in determining the effects of HNF-3 on cell-specific CCSP gene expression in the bronchiolar epithelium.
机译：克拉拉细胞分泌蛋白（CCSP）基因的5'侧翼区域包含两个顺式作用元件，它们在体外与肝细胞核因子（HNF）-3α和HNF-3β结合。为了确定这些蛋白质在支气管上皮中介导CCSP基因表达中的作用，将含有CCSP 5'侧翼区域各个区域的嵌合CCSP-reporter基因构建体与HNF-3 alpha或HNF-共转染到H-441细胞中3个beta表达质粒。这些研究表明，这些转录因子均能积极调节CCSP基因的表达，并揭示CCSP I区（-132至-76）足以介导此作用。使用对应于CCSP区I的寡核苷酸，细支气管上皮细胞的核提取物和HNF-3特异性抗体进行的凝胶迁移试验，表明HNF-3 alpha和HNF-3 beta是支气管上皮细胞中唯一与之直接相互作用的蛋白这个地区。与这些观察结果一致，发现HNF-3α和HNF-3β转录物在该细胞群中富集，并且成年肺的原位杂交揭示了在表达CCSP基因的非纤毛细支气管上皮细胞中HNF-3基因表达。最后，使用CCSP区域I和异源启动子进行的实验表明，该区域在启动子特异性背景下起作用，这表明通过最小CCSP启动子区域相互作用的其他因素对于确定HNF-3对细胞特异性CCSP基因的作用至关重要在支气管上皮中表达

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