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，可导致危及生命的全身感染，尤其是在免疫功能低下的个体中。靶向Hsp 90分子伴侣蛋白为真菌疾病提供了强有力的治疗策略。然而，临床实用性取决于鉴定Hsp 90网络的组分，其可以在病原体中选择性地靶向而不伤害受感染的宿主。 蛋白质组学和化学基因组学方法的结合将提供对C.白色念珠菌这将检验Hsp 90及其辅伴侣在特定环境条件下与不同的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Hsp 90是蛋白质稳态的中心枢纽，拟议的研究将确定Hsp 90相互作用与应激反应，耐药性，形态发生和毒力的重要作用。将根据以下因素对这些相互作用物进行优先排序：1）在多个屏幕中识别; 2） 遗传相互作用或突变表型; 3）相互作用的新奇。所有物理相互作用将通过相互免疫共沉淀进行验证，所有遗传相互作用或形态发生缺陷将通过与野生型等位基因互补进行验证。上位性分析将确定Hsp 90遗传网络的结构。最后，将评价候选靶标在毒力中的作用。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控制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.