The nucleolus as a therapeutic target for A fumigatus

性状（由申请方提供）：烟曲霉是一种 一种机会性真菌病原体，可引起严重的，通常是致命的感染， 免疫力低下的患者。虽然侵入性的发病率 曲霉病继续上升，反映了人口的增加， 免疫抑制患者，只有少数抗真菌药物可用于 治疗和它们的功效受到毒性问题的严重阻碍， 耐药性由于大多数患者会死于曲霉菌病， 接受治疗后，迫切需要开发新药， 治疗这种疾病。在这项资助中，我们提出核仁蛋白质是 新的治疗靶点的重要考虑因素，因为它们的有效 对蛋白质合成的影响我们的初步数据显示， 真菌特异性核仁蛋白，对S. cerevisiae，我们建议测试的假设，A.烟曲霉 直系同源物AfcgrA定义了核仁功能的途径， 有效治疗新靶点。 目的I的目标是使用A.烟曲霉模型 一种假定的核仁靶向药物的治疗效果。Afcgra 基因将被基因靶向方法破坏， 突变的等位基因损害疾病进展的速度或组织的模式 将在曲霉病的两种体内模型中比较传播。 Aim II的目标是获取关于AfcgrA相互作用的信息， 促进未来基于结构的药物设计应用于 AfcgrA抑制剂的鉴定。我们将使用酵母双杂交系统， 确定AfcgrA的分子伴侣，并深入了解结构 通过确定这些交互的域组织， AfcgrA与圆二色性光谱。 Aim III的目标是确定AfcgrA的区域， 核仁定位我们建议定义这些序列，预计， 可以利用对用于定位AfcgrA的机制的理解， 这种转运过程的选择性抑制剂的设计。

DESCRIPTION (provided by the applicant): Aspergillus fumigatus is an opportunistic fungal pathogen that causes severe and often fatal infections in patients with depressed immunity. Although the incidence of invasive aspergillosis is continuing to rise, reflecting the increased population of immunosuppressed patients, there are only a few antifungal agents available for treatment and their efficacy is severely hampered by problems with toxicity and drug resistance. Since most patients will die from aspergillosis despite having received treatment, there is an urgent need for the development of new drugs to treat this disease. In this grant we are proposing that nucleolar proteins are important considerations for novel therapeutic targets because of their potent effects on protein synthesis. Our preliminary data has identified a conserved fungal-specific nucleolar protein that is essential for growth in S. cerevisiae, and we are proposing to test the hypothesis that the A. fumigatus ortholog, AfcgrA, defines a pathway of nucleolar function that would be an effective new target for therapy. The goal of Aim I is to use AfcgrA-deficient mutants of A. fumigatus to model the therapeutic efficacy of a putative nucleolus-targeting drug. The AfcgrA gene will be disrupted by gene targeting approaches and the extent to which the mutant alleles impair the rate of disease progression or pattern of tissue dissemination will be compared in two in vivo models of aspergillosis. The goal of Aim II is to acquire information on AfcgrA interactions that will facilitate future application of structure-based drug design to the identification of AfcgrA inhibitors. We will use the yeast two-hybrid system to identify the molecular partners of AfcgrA, and gain insight into the structural requirements for these interactions by determining the domain organization of AfcgrA with circular dichroism spectroscopy. The goal of Aim III is to determine the regions of AfcgrA that specify nucleolar localization. We propose to define these sequences, anticipating that an understanding of the mechanism used to localize AfcgrA could be exploited in the design of selective inhibitors of this transport process.