Control of stage differentiation of Trypanosoma brucei by the host environment: a large scale RNAi screen of the kinome and phosphoproteomic analysis

宿主环境对布氏锥虫阶段分化的控制：大规模 RNAi 激酶组筛选和磷酸化蛋白质组分析

• 批准号: 63116995
• 负责人: Professor Dr. Michael Boshart
• 金额: --
• 依托单位: Institut für Genetik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2008
• 资助国家: 德国
• 起止时间: 2007-12-31 至 2012-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/63116995?language=en
• 关键词: Control; stage; differentiation; Trypanosoma; brucei

Trypanosomes shuttle between vertebrates and a blood feeding insect, the tsetse in tropical Africa. In the parasitic life cycle, rapid adaptation to sudden changes of the host environments are essential for survival. Signals provided by the host environment trigger coordinate changes in parasite gene expression that result in differentiation to adapted developmental stages. Our previous work shows that coincidence of temperature change (cold shock), citrate or cisaconitate in the medium, and cell cycle arrest is important to initiate differentiation to the fly midgut stage. We propose to identify molecular components of a yet unexplored pathway that transmits the host cues temperature and citrate. Two complementary approaches will be pursued: (1) screening of a bioinformatically identified set of 207 candidate genes, including all protein kinases in the T. brucei genome, using RNAi in a culture model of induced differentiation; (2) survey of protein phosphorylation changes induced by the host cues in 2D protein electrophoresis, and identification of additional pathway components by phosphoproteomics; (3) characterization of an already discovered cold-induced protein kinase. We expect unusual and novel signaling proteins, given the early phylogenetic branching of trypanosomes. Consequently, insights into the evolution of signaling and interorganismic control mechanisms, and discovery of potential drug targets for presently insufficient therapy of important tropical diseases may result from this project.

锥虫在脊椎动物和热带非洲的吸血昆虫采采蝇之间穿梭。在寄生生命周期中，快速适应宿主环境的突然变化对于生存至关重要。宿主环境提供的信号触发寄生虫基因表达的协调变化，从而导致分化到适应的发育阶段。我们之前的工作表明，温度变化（冷激）、培养基中的柠檬酸盐或顺乌头酸盐以及细胞周期停滞的同时发生对于启动向果蝇中肠阶段的分化非常重要。我们建议鉴定一种尚未探索的传递宿主线索温度和柠檬酸盐途径的分子成分。将采用两种互补的方法：(1) 在诱导分化培养模型中使用 RNAi 筛选一组生物信息学鉴定的 207 个候选基因，包括 T. brucei 基因组中的所有蛋白激酶； (2) 调查二维蛋白质电泳中宿主线索诱导的蛋白质磷酸化变化，并通过磷酸蛋白质组学鉴定其他途径成分； (3) 已发现的冷诱导蛋白激酶的表征。鉴于锥虫的早期系统发育分支，我们期待不寻常和新颖的信号蛋白。因此，该项目可能会深入了解信号传导和生物体间控制机制的演变，并发现目前对重要热带疾病治疗不足的潜在药物靶点。

项目成果

Adenylate Cyclases of Trypanosoma brucei Inhibit the Innate Immune Response of the Host

• DOI: 10.1126/science.1222753
• 发表时间: 2012-07-27
• 期刊: SCIENCE
• 影响因子: 56.9
• 作者: Salmon, Didier;Vanwalleghem, Gilles;Pays, Etienne
• 通讯作者: Pays, Etienne

Cytokinesis of Trypanosoma brucei bloodstream forms depends on expression of adenylyl cyclases of the ESAG4 or ESAG4‐like subfamily

• DOI: 10.1111/j.1365-2958.2012.08013.x
• 发表时间: 2012-04
• 期刊: Molecular Microbiology
• 影响因子: 3.6
• 作者: D. Salmon;S. Bachmaier;Carsten Krumbholz;Markus Kador;J. Gossmann;P. Uzureau;E. Pays;M. Boshart