CYTOSKELETAL AND SIGNALING MECHANISMS REGULATING CILIARY TRAFFIC

调节纤毛交通的细胞骨架和信号传导机制

• 批准号: 10207107
• 负责人: PRACHEE AVASTHI
• 金额: $ 41万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10207107
• 关键词: Actins; Affect; Apical; Bardet-Biedl Syndrome; Biological Assay; Biological Models; Cell surface; Cellular biology; Chlamydomonas reinhardtii; Destinations; Disease; Eukaryota; Functional disorder; Genes; Genetic Screening; Goals; Leber' s amaurosis; Microtubules; Modeling; Molecular; Molecular Motors; Motor; Myosin ATPase; Organelles; Pathogenesis; Pathway interactions; Phosphoric Monoester Hydrolases; Phosphotransferases; Play; Polycystic Kidney Diseases; Primary Ciliary Dyskinesias; Protein Biosynthesis; Proteins; Proteomics; Regulation; Role; Signal Pathway; Signal Transduction; Situs Inversus; Work; base; body system; chemical genetics; ciliopathy; new therapeutic target; novel; protein transport; trafficking

Project Summary/Abstract A fundamental question in cell biology is how targeted intracellular protein trafficking is achieved and regulated. An excellent framework to ask this question is to study transport to a specific organelle or intracellular compartment. Trafficking of proteins to and into the eukaryotic flagellum is an ideal model to study polarized transport given that flagellar protein synthesis and trafficking can be induced experimentally on-demand, cargo proteins have been identified through proteomics and the ultimate cargo destination is localized to a very small region at the apical cell surface. This trafficking pathway was previously thought to only require microtubules and the regulation of microtubule motors through signaling pathways. Through quantitative analysis of flagellar motor dynamics in the canonical flagellar model system Chlamydomonas reinhardtii, we discovered that actin and an actin-based myosin motor play an important role in regulating the localization and compartmentalization of flagellar proteins. We also identified a variety of signaling pathways including a phosphatase, MKP-2, that are required for proper flagellar assembly. The broad goals of our work are to: 1) use chemical and genetic screening to identify novel pathways that integrate to control flagellar protein trafficking and molecular motors flagellar entry; and 2) use a toolbox of cellular and molecular assays to dissect the mechanisms by which they exert this control. We expect to uncover entirely new avenues for the study of secretory pathways conserved in all eukaryotes as well as novel functions for known genes in coordinated cellular trafficking.

项目摘要/摘要 细胞生物学的一个基本问题是如何实现靶向细胞内蛋白质运输以及 受监管。问这个问题的一个绝佳框架是研究运输到特定细胞器或细胞内的运输 车厢。将蛋白质运输到真核鞭毛中是研究极化的理想模型 鉴于鞭毛蛋白的合成和运输可以通过实验诱导货物的运输 蛋白质已经通过蛋白质组学鉴定出蛋白质，最终的货物目的地位于非常小的 顶端细胞表面的区域。以前认为这种贩运途径仅需要微管 以及微管电动机通过信号通路的调节。通过鞭毛的定量分析 典型的鞭毛模型系统中的电动机动力学Chlamydomonas renhardtii，我们发现肌动蛋白和肌动蛋白和 基于肌动蛋白的肌球蛋白运动在调节定位和分隔中起着重要作用 鞭毛蛋白。我们还确定了各种信号通路，包括磷酸酶MKP-2， 适当的鞭毛组件所需。我们工作的广泛目标是：1）使用化学和遗传筛查 确定整合以控制鞭毛蛋白运输和分子电动机鞭毛的新途径 入口; 2）使用细胞和分子测定的工具箱来剖定它们施加的机制 控制。我们希望发现全新的分泌途径的全新途径 真核生物以及已知基因在协调的细胞运输中的新功能。