RNA-Protein Interactions in Flavivirus Infection

描述（由申请人提供）：黄热病（YF）由YF病毒（YFV）引起，每年约有200，000人患病，主要发生在撒哈拉以南非洲地区，在南美洲热带地区也有少量发生。YF被认为是一个重新出现的威胁，特别令人关切的是，南美洲和中美洲已经成熟，未来可能发生毁灭性的流行病。登革热（DF）是由四种登革热病毒（DENs）之一引起的，是热带和亚热带国家的一个主要和复发的健康问题。对处于危险中的人口的保守估计是惊人的25亿人。DF可发展为登革出血热（DHF）和登革休克综合征（DSS），这是可怕的发展。所有这些考虑因素共同使青年和糖尿病成为主要的和日益严重的世界健康问题。此外，这些病毒对美国人口构成威胁，最近全国爆发的相关西尼罗河病毒就是明证。该申请解决的长期问题是，由YF和DF（以及其他黄病毒）引起的疾病目前无法治愈。因此，开发针对这两种病毒的新疗法非常重要。该应用的首要目标是通过鉴定宿主蛋白质和病毒RNA之间的新型相互作用来发现新的潜在治疗靶点。为了实现这一目标，我们提出了以下具体目标：1）使用RNA亲和层析和蛋白质组学分析，以确定宿主蛋白质，特异性地与重要的病毒RNA元件相互作用。这一目标将需要实验确认RNA的结构和元素，构建和测试的RNA亲和矩阵，和蛋白质组学分析的特定相互作用的蛋白质。2)验证所鉴定的RNA结合蛋白的功能重要性。我们将使用分子遗传学和RNA干扰来确定，在病毒RNA结合蛋白，YFV和DEN繁殖所需的因素。这个探索性项目的成功完成将使我们能够开发一套假设驱动的研究，以解决这些病毒RNA-宿主蛋白相互作用的分子细节及其作为治疗靶点的可行性。本申请所解决的长期公共卫生问题是由黄热病和登革热病毒以及相关病毒（例如，西尼罗河热病毒）目前没有治愈方法。该应用的总体目标是通过鉴定病毒（RNA）组分与细胞因子之间的新型相互作用来发现新的潜在治疗靶点。我们希望这项工作将导致新的抗病毒疗法的发展。

DESCRIPTION (provided by applicant): Yellow Fever (YF), which is caused by YF virus (YFV), afflicts about 200,000 individuals yearly mostly in sub-Saharan Africa and to a lesser extent in the tropical regions of South America. YF is considered a reemerging threat and there is particular concern that South and Central America are ripe for future devastating epidemics. Dengue Fever (DF), which is caused by one of four dengue viruses (DENs), is a major and resurging health problem in tropical and sub-tropical countries. A conservative estimate of the population at risk is a staggering 2.5 billion people. DF can progress to dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS), which are dreaded developments. All of these considerations conspire to make YF and DF, major and growing world health problems. Additionally these viruses represent a threat to the U.S. population, something made evident by the recent national outbreak of the related West Nile virus. The long-term problem addressed by this application is the fact that the illnesses caused by YF and DF (and other flaviviruses) currently have no cure. It is important, therefore, to develop novel therapeutics for these two viruses. The overarching goal of this application is to discover new potential therapeutic targets by identifying novel interactions between host proteins and viral RNAs. In order to accomplish this we propose the following specific aims: 1) To use RNA affinity chromatography and proteomic analysis to identify host proteins that specifically interact with important viral RNA elements. This aim will require experimental confirmation of RNA structures and elements, construction and testing of RNA affinity matrices, and proteomic analysis of specific interacting proteins. 2) To validate the functional importance of the identified RNA binding proteins. We will use molecular genetics and RNA interference to identify, among the viral RNA binding proteins, the factors required for YFV and DEN propagation. Successful completion of this exploratory project will permit us to develop a hypothesis-driven set of investigations to address the molecular details of these viral RNA-host protein interactions and their feasibility as therapeutic targets. The long-term public health problem addressed by this application is the fact that the illnesses caused by Yellow Fever and Dengue Fever viruses, and related viruses (e.g., West Nile Fever virus) currently have no cure. The overarching goal of this application is to discover new potential therapeutic targets by identifying novel interactions between components of the virus (RNA) and cellular factors. We hope this work will lead to the development of new antiviral therapies.