RNA-Protein Interactions in Flavivirus Infection

描述(申请人提供)：黄热病(YF)，由YF病毒(YFV)引起，每年困扰大约20万人，主要发生在撒哈拉以南非洲，在南美洲的热带地区程度较小。YF被认为是一个新出现的威胁，尤其令人担忧的是，南美洲和中美洲的未来破坏性流行病的时机已经成熟。登革热是由四种登革热病毒之一引起的，在热带和亚热带国家是一个主要的和死灰复燃的健康问题。据保守估计，面临风险的人口有25亿人，令人震惊。登革热可发展为登革出血热(DHF)和登革休克综合征(DSS)，这些都是令人恐惧的发展。所有这些考虑共同导致YF和DF成为日益严重的世界卫生问题。此外，这些病毒对美国人口构成威胁，最近在全国范围内爆发的相关西尼罗河病毒就证明了这一点。这个应用程序解决的长期问题是，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是很重要的。这项应用的首要目标是通过识别宿主蛋白和病毒RNA之间的新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)利用RNA亲和层析和蛋白质组学分析来鉴定与重要病毒RNA元件特异相互作用的宿主蛋白。这一目标将需要对RNA结构和元件进行实验确认，构建和测试RNA亲和力矩阵，并对特定相互作用的蛋白质进行蛋白质组学分析。2)验证已鉴定的RNA结合蛋白的功能重要性。我们将利用分子遗传学和RNA干扰在病毒RNA结合蛋白中鉴定YFV和DEN繁殖所需的因子。这一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。该应用程序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(例如西尼罗热病毒)引起的疾病目前无法治愈。这项应用的首要目标是通过识别病毒成分(RNA)和细胞因子之间的新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开发。

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.