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.

项目概要/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并 受监管。提出这个问题的一个很好的框架是研究特定细胞器或细胞内的运输 隔间。蛋白质进出真核鞭毛的运输是研究极化的理想模型 鉴于鞭毛蛋白合成和运输可以通过实验按需诱导，货物运输 蛋白质已经通过蛋白质组学鉴定出来，最终的货物目的地局限于一个非常小的区域。 位于顶端细胞表面的区域。这种运输途径以前被认为只需要微管 以及通过信号通路调节微管马达。通过对鞭毛的定量分析 在典型的鞭毛模型系统莱茵衣藻的运动动力学中，我们发现肌动蛋白和 基于肌动蛋白的肌球蛋白运动在调节肌球蛋白的定位和区划方面发挥着重要作用 鞭毛蛋白。我们还确定了多种信号传导途径，包括磷酸酶 MKP-2， 正确的鞭毛组装所必需的。我们工作的主要目标是：1）使用化学和基因筛查 确定整合控制鞭毛蛋白运输和鞭毛分子马达的新途径 入口; 2）使用细胞和分子检测工具箱来剖析它们发挥这种作用的机制 控制。我们期望为研究所有生物中保守的分泌途径找到全新的途径 真核生物以及协调细胞运输中已知基因的新功能。