RUI: Probing Intracellular Trafficking Decisions using Quantitative Fluorescence Microscopy and Biochemical Methods

所有真核细胞都必须根据生理条件做出转运决定，以维持跨膜或膜相关蛋白的最佳分布。虽然已经描述了许多信号通路和基本的贩运蛋白质，但关于如何作出贩运决定的信息有限。本项目将研究铜代谢基因murr1结构域蛋白在铜转运调控中的作用，这是目前转运研究领域的热点。这项研究的结果对了解其他膜蛋白的转运具有潜在的重要意义。该项目的社会影响是教育，旨在在阿拉斯加-安克雷奇大学（UAA）建立分子和细胞生物科学的培训机会，这方面有很高的需求，但能力有限。该项目将在UAA建立一个综合研究和教育计划，并增加本科生在研究方面的培训机会。本科生将直接参与发现过程，同时获得包括定量生物科学在内的先进科学概念的经验。本课题的研究成果将直接融入探究型细胞生物学课程的课堂活动中。所有真核细胞必须平衡膜蛋白和脂质向质膜的流出运输和向高尔基体的逆行运输。虽然已经研究了许多信号通路和必需的转运蛋白，但关于细胞如何整合各种信号以做出逆行转运决策的信息尚未阐明。COMMD1（铜代谢基因murr1结构域）是影响参与钠、氯和铜运输的膜蛋白的运输和稳定性的蛋白家族的创始成员。铜是维持正常细胞功能的特定酶的重要组成部分。本项目旨在了解COMMD1作为接头蛋白在铜atp酶转运中的作用，铜atp酶在铜转运中起重要作用。将采用一系列定量生化和生物物理技术，包括荧光显微镜、共定位研究和蛋白质3D结构的分子建模。这些结果对于理解其他膜蛋白的逆行转运具有潜在的重要意义。

All eukaryotic cells must make trafficking decisions to maintain optimal distribution of transmembrane or membrane-associated proteins in response to physiological conditions. While a number of signaling pathways and essential trafficking proteins have been described, the information on how trafficking decisions are made is limited. This project will study an area of great interest in the trafficking field, which is the role of copper metabolism gene murr1 domain protein in regulating copper transport. The results of this study are potentially significant for understanding trafficking of other membrane proteins. The societal impacts of this project are educational, and aim to build training opportunities in molecular and cellular biosciences at University of Alaska-Anchorage (UAA), for which there is a high demand, yet limited capacity. This project will build an integrated research and education program at UAA and enhance training opportunities for undergraduate students in research. Undergraduate students will participate directly in the discovery process while gaining experience with advanced scientific concepts, including quantitative biological sciences. The research results of this project will be directly integrated into classroom activities in an inquiry-based cell biology course.All eukaryotic cells must balance outflow trafficking of membrane proteins and lipids towards the plasma membrane with retrograde trafficking towards the Golgi. While a number of signaling pathways and essential trafficking proteins have been studied, the information on how cells integrate diverse signals to make retrograde trafficking decisions is yet to be elucidated. COMMD1 (copper metabolism gene murr1 domain) is the founding member of a family of proteins that influences trafficking and stability of membrane proteins involved in sodium, chloride, and copper transport. Copper is an important part of specific enzymes that maintain normal cell functions. This project aims to understand the role of COMMD1 as an adaptor protein in regulating trafficking of a copper ATPase, which has an important role in copper transport. A set of quantitative biochemical and biophysical techniques, including fluorescence microscopy, colocalization studies, and molecular modeling of protein 3D structures will be employed. These results are potentially significant for understanding the retrograde trafficking of other membrane proteins.