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

所有的真核细胞都必须做出运输决定，以维持跨膜或膜相关蛋白在生理条件下的最佳分布。虽然已经描述了一些信号通路和必要的贩运蛋白，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结构域蛋白在调控铜转运中的作用。本研究的结果对于理解其他膜蛋白的运输具有潜在的意义。该项目的社会影响是教育性的，旨在在阿拉斯加大学安克雷奇分校（UAA）建立分子和细胞生物科学的培训机会，这是一个高需求，但能力有限。该项目将在UAA建立一个综合的研究和教育计划，并提高研究本科生的培训机会。本科生将直接参与发现过程，同时获得先进的科学概念，包括定量生物科学的经验。本项目的研究成果将直接融入到探究式细胞生物学课程的课堂活动中。所有真核细胞都必须平衡膜蛋白和脂质向质膜的外流运输与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运输蛋白，但关于细胞如何整合不同信号以做出逆行运输决定的信息尚未阐明。COMMD1（铜代谢基因murr1结构域）是影响参与钠、氯和铜转运的膜蛋白的运输和稳定性的蛋白质家族的创始成员。铜是维持正常细胞功能的特定酶的重要组成部分。该项目旨在了解COMMD 1作为衔接蛋白在调节铜ATP酶运输中的作用，铜ATP酶在铜运输中具有重要作用。一套定量的生物化学和生物物理技术，包括荧光显微镜，共定位研究，和蛋白质三维结构的分子建模将被采用。这些结果对于理解其他膜蛋白的逆行运输具有潜在的意义。

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.