Roles of endosomal phosphoinositides in intracellular trafficking

内体磷酸肌醇在细胞内运输中的作用

• 批准号: RGPIN-2016-06155
• 负责人: Côté, Marceline
• 金额: $ 2.26万
• 依托单位: University of Ottawa
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2016
• 资助国家: 加拿大
• 起止时间: 2016-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=616133
• 关键词: Roles; endosomal; phosphoinositides; intracellular; trafficking

Endolysosomal trafficking is a tightly regulated process that involves membrane fusion and fission events, which ensure proper delivery and exchange of cargoes within endosomes, lysosomes, and other intracellular organelles or vesicles. It is essential for several cellular functions such as protein and lipid transport, autophagy, and degradation of macromolecules in lysosomes. How it is regulated is a fundamental issue in cellular biology. Our research program intends to use viruses as means to monitor and understand vesicular trafficking in cells. Indeed, viruses have evolved mechanisms to use and even regulate endosomal maturation or endolysosomal trafficking, since it can determine whether the infection is successful or not. Therefore, they can represent powerful and invaluable tools for the study of cellular trafficking pathways. We found that proteins of the Vac14/Fig4/PIKfyve complex, which regulates the synthesis of the phosphoinositide(3,5)bisphosphate at the endolysosomal membrane, are essential for infection by Ebola virus (EboV) and severe acute respiratory syndrome coronavirus (SARS-CoV). How the enzymatic complex synthesizing PI(3,5)P2 is activated and the role of PI(3,5)P2 in endolysosomal trafficking remain unclear. Here, we propose to use EboV and SARS-CoV to answer these questions. The goal of this research program is to understand how this complex and its resulting phosphoinositide are regulated and how they control endolysosomal trafficking in cells, a fundamental cellular process. Hypothesis: The endosomal phosphoinositide PI(3,5)P2 is critical for the trafficking of viral particles to intracellular compartments where viral fusion occurs. Short-term objectives: 1) Examine the effect of genetic and chemical manipulation of PI(3,5)P2 levels 2) Perform a molecular dissection of the Vac14/Fig4/PIKfyve complex 3) Identify PI(3,5)P2 effectors Our long term goal is to gain a molecular understanding of the Vac14/Fig4/PIKfyve interactome and function. To meet our objectives, we will use a combination of genetic, chemical biology, and virological tools. Combining these specific tools with our expertise in molecular/cell biology and viral entry, we hope to fill a gap in the understanding of endolysosomal trafficking and how this fundamental process can be hijacked by viral pathogens.

内侧溶血贩运是一个严格调节的过程，涉及膜融合和裂变事件，可确保在内体，溶酶体和其他细胞内细胞器或囊泡内的适当输送和交换。对于多种细胞功能，例如蛋白质和脂质转运，自噬和大分子溶酶体中大分子的降解至关重要。如何调节是细胞生物学中的一个基本问题。我们的研究计划打算使用病毒作为监测和理解细胞中囊泡运输的手段。实际上，病毒已经发展了用于使用的机制，甚至调节内体成熟或内溶血贩运，因为它可以确定感染是否成功。因此，它们可以代表研究细胞运输途径的强大而有价值的工具。我们发现，VAC14/FIG4/PIKFYVE复合物的蛋白质调节内溶性肌体膜上的磷酸肌醇（3,5）双磷酸的合成对于通过埃博拉病毒病毒（EBOV）和严重的急性急性爆炸性综合征综合征Coronavirus（Sars-Cov）感染至关重要。如何激活酶复合物合成PI（3,5）P2，PI（3,5）P2在内溶性溶血体运输中的作用尚不清楚。在这里，我们建议使用EBOV和SARS-COV回答这些问题。 该研究计划的目的是了解该复合物及其产生的磷酸肌醇如何受到调节，以及它们如何控制细胞中的内溶液性贩运，这是一种基本的细胞过程。 假设： 内体磷酸肌醇Pi（3,5）P2对于将病毒融合的细胞内室进行运输至关重要。 短期目标： 1）检查PI的遗传和化学操作的影响（3,5）P2水平 2）对VAC14/FIG4/PIKFYVE复合物进行分子解剖 3）识别PI（3,5）P2效应子 我们的长期目标是对VAC14/FIG4/PIKFYVE ITSOTSPOTOME和功能的分子理解。 为了实现我们的目标，我们将结合遗传，化学生物学和病毒学工具。将这些特定工​​具与分子/细胞生物学和病毒进入方面的专业知识相结合，我们希望填补对内溶性贩运的理解以及如何通过病毒病原体劫持这种基本过程的空白。