Global analysis of Hsp90 client proteins in Candida albicans

 描述(由申请人提供)：白色念珠菌是一种主要的人类真菌病原体，可导致危及生命的系统性感染，特别是在免疫功能低下的个人中。靶向Hsp90伴侣蛋白为真菌疾病提供了一种强大的治疗策略。然而，临床用途取决于识别Hsp90网络的组件，这些组件可以选择性地在病原体中靶向，而不会伤害受感染的宿主。蛋白质组学和化学基因组学方法的结合将首次对白色念珠菌中的Hsp90伴侣网络进行全球分析。这将检验Hsp90及其辅助伴侣在特定环境条件下与不同客户蛋白相互作用的假设，从而实现一系列允许毒力的适应性反应。由于Hsp90是蛋白质动态平衡的中枢，这项拟议的研究将确定在应激反应、耐药性、形态发生和毒力方面具有重要作用的Hsp90相互作用因子。这些交互作用的优先顺序将基于：1)在多个屏幕上识别；2)大小和重复性 遗传互作或突变表型；3)互作的新颖性。所有的物理相互作用将通过相互免疫共沉淀来验证，所有的遗传相互作用或形态发生缺陷将通过与野生型等位基因的互补来验证。上位性分析将确定Hsp90遗传网络结构。最后，将评估候选靶标在毒力方面的作用。这项工作将揭示抗真菌治疗的新靶点，并阐明最古老和最保守的细胞调节因子之一管理真菌生物学和疾病的机制。

 DESCRIPTION (provided by applicant): Candida albicans is a leading human fungal pathogen that causes life-threatening 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 target for antifungal therapeutics and illuminate the mechanisms by which one of the most ancient and conserved cellular regulators governs fungal biology and disease.