Location: Its role in protein-RNA contact during trypanosome gene regulation

描述(申请人提供)：锥虫是一种寄生虫，会在人和牲畜中引起衰弱的、往往是致命的疾病。在撒哈拉以南非洲，一个特别严重的问题是由布鲁氏锥虫引起的，这是一种寄生虫，通过受感染的采采蝇的叮咬传播。布鲁氏毛滴虫通过在其哺乳动物宿主的血液、组织空间以及最终的中枢神经系统中繁殖而致病。相关寄生虫对世界其他地区的人类种群同样重要，包括南美洲和中美洲、东南亚和中东。我们的长期目标是在生化和遗传细节上了解锥虫基因表达的协调机制。为了实现这一目标，我们正在专门研究寄生虫用来调节其mRNA表达模式的机制。信使核糖核酸的表达决定了寄生虫的代谢能力，形成了一个特有的必需蛋白质组。作为理解mRNA表达调控的途径，我们正在研究一种新的RNA结合蛋白RBP42，之所以选择它进行研究，是因为它(1)是寄生虫增殖所必需的，(2)与细胞能量代谢途径中编码各种酶和蛋白质的mRNAs编码区(开放阅读框)结合。我们目前还不知道其他锥虫蛋白具有这种结合特异性；然而，在神经生物学和发育领域，人们对编码区结合蛋白的认识越来越多。此外，我们的数据表明，RBP42是mRNA翻译、mRNA稳定性或mRNA隔离的重要因素，因为它在布鲁氏毛滴虫生活史的前周期阶段与多聚体相关的翻译mRNAs结合。在我们的目标中，我们将通过确定RBP42在布鲁氏毛滴虫基因表达网络中的作用和作用方式来探讨RBP42的功能。这个探索性的、为期两年的项目基于我们实验室最近发表的初步数据、初步数据和补充的体内分子遗传学和体外生化方法。

DESCRIPTION (provided by applicant): Trypanosomatids are parasites that cause debilitating and often fatal diseases in humans and livestock. A particularly serious problem in sub-Saharan Africa is caused by Trypanosoma brucei, a parasite spread by the bite of an infected Tsetse fly. T. brucei causes disease by proliferating in the blood, tissue spaces, and eventually the central nervous system of its mammalian host. Related parasites are equally important to human populations in other parts of the world, including South and Central America, Southeastern Asia and the Middle East. Our long-term goal is to understand, in biochemical and genetic detail, the mechanisms that coordinate gene expression in trypanosomatids. To achieve this goal, we are specifically investigating the mechanism used by the parasites to regulate their mRNA expression pattern. mRNA expression gives rise to a characteristic and essential proteome and determines the metabolic capabilities of the parasites. As a pathway into understanding mRNA expression control, we are studying a novel RNA-binding protein, RBP42, which was chosen for investigation because it (1) is essential for parasite proliferation and (2) binds to the coding region (open reading frame) of mRNAs encoding a variety of enzymes and proteins within the cells' energy metabolic pathways. We currently know of no other trypanosome protein with this binding specificity; however there is an increasing awareness of coding region binding proteins in the fields of neurobiology and development. Furthermore, our data suggest that RBP42 is an important factor in mRNA translation, mRNA stability or mRNA sequestration as it binds to polysome-associated, translating mRNAs during the procyclic stage of the T. brucei life-cycle. In our aim, we will explore the function of RBP42 by determining its role and mode of action in gene expression networks of T. brucei. This exploratory, two-year project is grounded in preliminary data, in a recent publication from our laboratory, preliminary data and complementary in vivo molecular genetic and in vitro biochemical approaches.