The nucleolus as a therapeutic target for A fumigatus

描述(申请人提供)：烟曲霉是一种 一种机会性真菌病原体，可导致严重的，通常是致命的感染 免疫力低下的患者。尽管侵袭性疾病的发生率 曲霉病继续上升，反映出 免疫抑制患者，只有少数几种抗真菌药物可用于 治疗和疗效严重受制于毒性和 抗药性。因为大多数患者都会死于曲霉菌病，尽管 接受治疗后，迫切需要开发新药来 治疗这种疾病。在这项资助中，我们提出核仁蛋白是 新的治疗靶点的重要考虑因素是它们的效力 对蛋白质合成的影响。我们的初步数据确定了一个保守的 真菌特有的核仁蛋白，对S. 我们建议检验一种假设，即烟曲霉菌 原基因AfcgrA定义了核仁功能的一条途径，这条途径将是一种 有效的治疗新靶点。 目标I的目标是使用缺乏AfcgrA的烟曲霉菌突变体来模拟 一种假定的核仁靶向药物的治疗效果。《非洲人》 基因将被基因打靶方法扰乱，以及基因打靶的程度 突变等位基因影响疾病进展速度或组织模式 在曲霉病的两个活体模型中，将对传播进行比较。 AIM II的目标是获得关于AfcgrA相互作用的信息， 促进基于结构的药物设计在未来的应用 AfcgrA抑制剂的鉴定。我们将使用酵母双杂交系统来 确定AfcgrA的分子伙伴，并深入了解其结构 通过确定域组织来满足这些交互的要求 具有圆二色光谱的AfcgrA。 目标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 对用于本地化AfcgrA的机制的理解可以在 这一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.