CCSP Promoter Function and Tumor Necrosis Factor

描述（由申请人提供）：本修订申请旨在阐明急性呼吸道感染和炎症期间肺特异性基因的转录调控机制。感染期间肺功能的稳态可能是由关键的肺特异性基因调控的。急性感染后，重要的肺内稳态蛋白，包括表面活性剂蛋白（SP）基因和Clara细胞分泌蛋白（CCSP）显著降低。本文的初步数据表明，CCSP基因表达通过启动子功能的衰减而下调，特别是在含有几个关键反激活结合位点的近端启动子区域。同样，新的发现暗示炎症介质，特别是tnf - α，在宿主对急性感染的反应中调节肺基因转录的作用。CCSP基因和SP基因的近端启动子元件存在很强的相似性，这表明这些基因的转录调控机制可能在正常和病变肺中都是协调的。利用CCSP转录调控作为模型，现在可以利用转基因方法在体外和体内系统中研究肺特异性基因的转录调控和dna -蛋白质相互作用。具体来说，该应用程序旨在描述CCSP启动子位点中的dna -蛋白质相互作用，并阐明在宿主对急性感染的反应中，通过特定炎症介质异常调节肺特异性基因表达的转录因子调控机制。使用CCSP启动子调控作为模型来理解小鼠肺中的转录调控具有明显的优势，包括能够开发转基因动物来研究肺特异性转录调控和体内dna -蛋白相互作用。利用这种方法，可以在体内评估病原体或炎症介质对肺特异性基因的转录调节机制；这一策略在其他转录调控基因的研究中通常是不可用的。阐明dna -蛋白相互作用和转录因子调控对肺特异性转录的减弱可能对理解严重呼吸道感染和炎症期间肺功能减弱的机制很重要。

DESCRIPTION (provided by applicant): This revised application seeks to elucidate the mechanisms of transcriptional regulation of lung-specific genes during acute respiratory infection and inflammation. Homeostasis of lung function during infection is likely regulated by critical lung-specific genes. Important lung homeostatic proteins, including surfactant protein (SP) genes and the Clara cell secretory protein (CCSP) are markedly decreased following acute infection. Preliminary data herein indicate that CCSP gene expression is down regulated through attenuation of promoter function, specifically in the proximal transactivating promoter region containing several critical transactivating binding sites. Likewise, new findings implicate the role of inflammatory mediators, in particular TNF-alpha, in the regulation of lung gene transcription during the host response to acute infection. Strong similarities exist in the proximal promoter elements of CCSP and SP genes, suggesting that mechanisms of transcriptional regulation of these genes may be coordinated both in normal and diseased lung. Using CCSP transcriptional regulation as a model, transcriptional regulation, and DNA-protein interactions of lung-specific genes can now be studied in both in vitro and in vivo systems using a transgenic approach. Specifically, this application proposes to delineate the DNA-protein interactions in the CCSP promoter loci and elucidate mechanisms of transcription factor regulation that aberrantly regulate lung-specific gene expression by specific inflammatory mediators during the host response to acute infection. The use of CCSP promoter regulation as a model for understanding transcriptional regulation in the mouse lung has distinct advantages, including the ability to develop transgenic animals for the study of lung-specific transcriptional regulation and DNA-protein interactions in vivo. Using this approach, the mechanisms of transcriptional regulation of lung-specific genes by pathogens or inflammatory mediators can be assessed in vivo; a strategy that is often unavailable for the study of other transcriptionally regulated genes. Elucidation of the DNA-protein interactions and transcription factor regulation that attenuate lung-specific transcription will likely be important to understand the mechanisms of diminished lung function during severe respiratory infections and inflammation.