Global analysis of Hsp90 client proteins in Candida albicans

摘要： 白色念珠菌是一种主要的人类真菌病原体，可导致生命危险。 全身性感染，尤其是免疫功能受损的个体。瞄准 Hsp90伴侣蛋白为真菌提供了一种有效的治疗策略 疾病。然而，临床应用取决于识别Hsp90的成分 可以选择性地在病原体中靶向而不会伤害感染者的网络 主持人。 蛋白质组学和化学基因组学方法的结合将提供第一个 白念珠菌Hsp90分子伴侣网络的全球分析。这将测试 Hsp90及其辅助伴侣与不同客户蛋白相互作用的假说 特定的环境条件，实现了一系列适应性响应，从而允许 致命性。 由于Hsp90是蛋白质动态平衡的中心枢纽，拟议中的研究将 确定在应激反应、耐药、 形态发生和致病力。将根据以下因素确定这些参与者的优先顺序：1) 多屏鉴定；2)基因的大小和重复性 相互作用或突变表型；以及3)相互作用的新颖性。全部为物理设备 相互作用将通过相互免疫共沉淀进行验证，所有基因 相互作用或形态形成缺陷将通过与 野生型等位基因。上位性分析将确定Hsp90基因的结构 网络。最后，将评估候选靶标在毒力方面的作用。这 这项工作将揭示抗真菌治疗的新靶点，并阐明其机制 一种最古老、最保守的细胞调节器通过它来控制真菌 生物学和疾病。

Summary: Candida albicans is a leading human fungal pathogen that causes lifethreatening systemic infections, especially in immunocompromised individuals. Targeting the Hsp90 chaperone protein provides a powerful therapeutic strategy for fungal disease. However, clinical utility depends upon identifying components of the Hsp90 network that can be selectively targeted in the pathogen without harming the infected host. A combination of proteomic and chemical genomic approaches will provide the first global analysis of the Hsp90 chaperone network in C. albicans. This will test the hypothesis that Hsp90 and its co-chaperones interact with different client proteins under specific environmental conditions, enabling a range of adaptive responses that allow for virulence. Since Hsp90 is a central hub for protein homeostasis, the proposed research will identify Hsp90 interactors with important roles in stress response, drug resistance, morphogenesis, and virulence. These interactors will be prioritized based on: 1) identification in multiple screens; 2) magnitude and reproducibility of the genetic interaction or mutant phenotype; and 3) novelty of the interaction. All physical interactions will be validated by reciprocal co-immunoprecipitation, and all genetic interactions or morphogenetic defects will be validated by complementation with the wild-type allele. Epistasis analysis will determine the structure of the Hsp90 genetic network. Finally, the candidate targets will be evaluated for their role in virulence. This work will reveal novel targets for antifungal therapeutics and illuminate the mechanisms by which one of the most ancient and conserved cellular regulators governs fungal biology and disease.