Hunting plunc: novel protein in mammalian upper airway
Hunting plunc: novel protein in mammalian upper airway
批准号:
6319392
负责人:
ELIZABETH E LECLAIR
金额:
$9.62万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
2001
资助国家:
美国
项目状态:
已结题
起止时间:
2001-04-20 至 2005-03-31
关键词:
cell differentiation embryo /fetus cell /tissue gene expression histogenesis in situ hybridization laboratory mouse messenger RNA physiologic stressor protein localization protein protein interaction protein purification protein sequence protein structure function respiratory epithelium secretory protein tissue /cell culture
中文摘要
描述(申请人摘要):包括植物在内的20多种生物,
细菌和果蝇的全部染色体内容都被破译了。通过
2003年,狗、老鼠和人类的全部基因组将会公之于众。世界上的生物学家
21世纪现在面临着解释这一基因的挑战
信息和分析特定的基因产品-个别蛋白质-是如何
调节组织和器官的特定功能。我们计划培养学生
研究人员将表征一个基因的表达和潜在的相互作用
新的小鼠基因产物,最近命名为PLANC。Plunc信使RNA是
在小鼠鼻腔受限区域异常高水平的表达,
气管和支气管上皮。Plunc本身似乎是一个秘密的
功能未知的蛋白质,有可能发生二聚化或
蛋白质之间的相互作用。因为上半身的呼吸道上皮
呼吸道是吸入空气首先遇到的器官,表现为受限
在该区域发现了一种新的蛋白质,这引发了这样一个问题:该蛋白质如何
有助于区分、维持或可能保护呼吸道。
最近,人类、大鼠和牛的部分序列已经被克隆,
提示PLANC蛋白在气管上皮中可能是保守的。
呼吸空气的生物体。尽管这种遗传结构的保守性和
组织特异性表达意味着蛋白质功能的保守性,很少是
已知PLEC在任何生命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何时何地在肺上皮细胞中表达
体内和体外的细胞。提出了三个具体目标:1)
观察PLANC在小鼠肺发育过程中的原位表达模式
其与上皮细胞分化的相关性,2)标记和跟踪
培养的肺细胞株分泌PANC蛋白,以及3)研究
Partic蛋白与a)执行蛋白的潜在相互作用
沉淀法观察PLANC是否与体内其他内源蛋白相互作用
肺环境,以及b)检查是否存在一过性的肺血管内皮细胞
当上皮细胞暴露于外源性应激源时受到调控,包括
毒素和发炎剂。综上所述,这些研究将提供一个
关于这种新的肺蛋白的丰富的基础科学信息。他们还将
为寻求经验的学生提供全面的培训计划
分子生物学、发育生物学和蛋白质生物化学。
英文摘要
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.
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