Thioredoxin localization and its function in the airway

硫氧还蛋白定位及其在气道中的功能

基本信息

批准号：
6933806
负责人：
Richart W Harper
金额：
$ 12.69万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-09-15 至 2008-03-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant) The primary objective of this research proposal is to identify how compartmentalization of thioredoxin (TRX) in the cell affects its function. The specific aims of this project are to identify where TRX localizes under conditions of oxidative stress, and how this localization affects gene regulation. The objectives of the grant proposal will be achieved in four distinct parts. First, we plan to show that TRX preferentially accumulates in the nucleus during oxidative stress. Second, we will manipulate the TRX gene in order to direct it to either localize in the nucleus or be secreted. We will construct TRX fusion plasmids that contain either TRX attached to a nuclear localization sequence or TRX attached to a secretory sequence. We will then compare differences in NF-kappa B- and AP-1 dependent promoter activity in cells overexpressing nuclear TRX compared with cells overexpressing secreted TRX. This will help establish the functional role of TRX compartmentalization. Third, we will establish that TRX localization is important for modulating interleukin-8 (IL-8) gene activity. Forth, using similar TRX expression plasmids as constructed for specific aim two, we will examine the effects of nucleus-directed TRX versus secreted TRX in mice exposed to ozone. Using this in vivo model, we plan to confirm the findings we obtained in our in vitro studies. The principal investigator's immediate research goals are to better understand the signaling mechanisms involved in airway epithelial cells in response to oxidative stress. Consistent with this objective, he currently has received funding for a three-year project examining the signaling mechanisms responsible for up-regulating IL-8 in airway epithelial cells exposed to cigarette smoke. It is the plan of the principle investigator to use this career development award as an excellent mechanism to transition into an independent investigator over the next five years. It is his hope that this grant funding will provide him with the protected research time needed in order to obtain the experience and publications needed to make him a competitive candidate for R01 funding.

描述（由申请人提供）该研究建议的主要目的是确定细胞中硫氧还蛋白（TRX）的分室化会影响其功能。该项目的具体目的是确定TRX在哪里定位氧化应激的条件以及该定位如何影响基因规定。赠款提案的目标将在四个中实现不同的部分。首先，我们计划表明TRX优先积累氧化应激期间的核。其次，我们将操纵TRX基因为了将其定位在原子核或分泌中。我们将构造包含附着在A上的TRX的TRX融合质粒核定位序列或附加到分泌序列的TRX。我们然后将比较NF-KAPPA B-和AP-1依赖启动子的差异与细胞相比，过表达核TRX的细胞活性过表达分泌的TRX。这将有助于确定 TRX隔室化。第三，我们将确定TRX本地化是对于调节白介素8（IL-8）基因活性很重要。 hort，使用类似的TRX表达质粒与针对特定目标二的构建相似，我们将检查核指导的TRX与分泌TRX在小鼠中的影响暴露于臭氧。使用此体内模型，我们计划确认发现结果我们在体外研究中获得了。主要调查员的即时研究目标是更好地了解涉及的信号机制气道上皮细胞响应氧化应激。与此一致目的，他目前已经为三年项目获得了资金检查负责上调IL-8的信号传导机制气道上皮细胞暴露于香烟烟雾中。这是主要研究者将该职业发展奖用作出色的职业发展奖在接下来的五个中过渡到独立研究者的机制年。他希望这笔赠款能为他提供为了获得经验而需要保护的研究时间出版物需要使他成为R01资金的竞争候选人。