Hunting plunc: novel protein in mammalian upper airway

描述（申请人摘要）：超过20种生物，包括植物， 细菌和果蝇的整个染色体内容都被解码了。通过 到2003年，整个狗、老鼠和人类的基因组将被发现。生物学家 世纪的科学家们现在面临着解释这种遗传的挑战。 信息和分析特定的基因产物-个别蛋白质- 调节组织和器官特异性功能。我们建议培养学生 研究人员描述表达和潜在的相互作用， 新的小鼠基因产物，最近命名为plunc。Plunc信使RNA是 在小鼠鼻的限制区域以异常高的水平表达， 气管和支气管上皮。Plunc本身似乎是一个秘密的 具有二聚化潜力的未知功能的蛋白质，或 蛋白质相互作用因为上呼吸道上皮 气道是吸入空气首先接触的器官， 在这个区域发现了一种新的蛋白质，这就提出了一个问题， 有助于分化、维持或可能保护气道。 最近，人类、大鼠和牛的部分序列已被克隆， 表明plunc蛋白可能在气管上皮中是保守的 呼吸空气的有机体。虽然这种遗传结构的保守性， 组织特异性表达意味着蛋白质功能的保守性， 了解plunc在任何生物中的发育或生理作用， 动物 研究该蛋白在哺乳动物呼吸系统中的功能 我们将分析这种蛋白质在肺上皮细胞表达的位置和时间， 在体内和体外的细胞。提出了三个具体目标：1） 观察plunc在小鼠肺发育过程中的原位表达模式， 它与上皮细胞分化的相关性，2）标记和跟踪 培养的肺细胞系中plunc蛋白的分泌，以及3）研究 plunc蛋白质的潜在相互作用通过a）执行蛋白质 沉淀，看看Plunc是否与其他内源性蛋白质相互作用， 肺环境，和B）观察plunc mRNA的水平是否在时间上是 当上皮细胞暴露于外源性应激源时， 毒素和炎性物质。这些研究将提供一个 关于这种新型肺蛋白的丰富的基础科学信息。他们还将 作为一个全面的培训计划，为学生寻求经验， 分子生物学、发育生物学和蛋白质生物化学。

DESCRIPTION (Applicant's Abstract): Over 20 organisms, including plants, bacteria, and fruit flies have had their entire chromosome contents decoded. By 2003, the entire dog, mouse and human genomes will be known. Biologists of the 21St century are now faced with the challenge of interpreting this genetic information and analyzing how specific gene products - the individual proteins- modulate tissue- and organ-specific functions. We propose to train student investigators to characterize the expression and potential interactions of a novel mouse gene product, recently named plunc. Plunc messenger RNA is expressed at unusually high levels in restricted areas of the mouse nasal, tracheal and bronchial epithelium. Plunc itself appears to be a secreted protein of unknown function with the potential for dimerization or protein-protein interactions. Because the respiratory epithelium of the upper airway is the first organ encountered by inhaled air, the restricted expression of a novel protein in this region raises the question of how the protein might contribute to differentiating, maintaining, or possibly protecting the airway. More recently, partial human, rat and bovine sequences have been cloned, indicating that the plunc protein may be conserved in the tracheal epithelium of air-breathing organisms. although this conservation of genetic structure and tissue-specific expression implies conservation of protein function, little is known about the developmental or physiological role of plunc in any living animal. To investigate the function of this protein in the mammalian respiratory system we will analyze where and when this protein is expressed by lung epithelial cells both in vivo and in vitro. Three specific aims are proposed: 1) to observe the in situ expression pattern of plunc in developing mouse lungs and its correlation with epithelial cell differentiation, 2) to tag and track the secretion of plunc protein in cultured lung cell lines, and 3) to investigate the potential interactions of plunc protein by a) performing protein precipitations to see if plunc interacts with other endogenous proteins in the lung environment, and b) to see if the level of plunc mRNA is temporally regulated when epithelial cells are exposed to exogenous stressors, including toxins and inflammatory agents. Taken together, these studies will provide a wealth of basic science information on this novel lung protein. They will also serve as a comprehensive training program for students seeking experience in molecular biology, developmental biology and protein biochemistry.