CAREER: Structural Studies of a Highly Conserved Dynein Light Chain and its Role in Dynein Assembly and Cargo Recruitment

职业：高度保守的动力蛋白轻链的结构研究及其在动力蛋白组装和货物招募中的作用

• 批准号: 0238094
• 负责人: Elisar Barbar
• 金额: $ 74.42万
• 依托单位: Ohio University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-15 至 2004-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0238094&HistoricalAwards=false
• 关键词: CAREER; Structural; Studies; Highly; Conserved

Cytoplasmic dynein is a motor protein complex that is essential in a number of processes including chromosome segregation during mitosis, movement of vesicular cargo, positioning of intracellular organelles, and the establishment of cell polarity in oocyte development. This project focuses on characterizing the role of dynein light chain LC8 from Drosophila in dynein assembly and cargo recognition. LC8 is highly conserved, with 94% sequence identity between Drosophila and human. The dynein intermediate chain IC74 mediates the interaction between the heavy and light chains, and the assembly of LC8 and IC74 is likely to be a key step in formation of a functional complex. To elucidate the basis for LC8/IC74 recognition, the structure of the complex will be determined using NMR spectroscopy. The swallow protein, Swa, an example of cargo, is an RNA binding protein, which is essential in establishing cell polarity in developing oocytes. The interaction of Swa with LC8 has been demonstrated in Drosophila. Identification of binding sites in this interaction will be determined by limited proteolysis and cross-linking studies.Broader Impact: Structural studies of LC8 and its interactions with IC74 and Swa will be a significant contribution to understanding both the assembly of the dynein motor complex and its attachment to cargo. This work will entail substantive contributions from a number of undergraduates. The project will also involve development of a class on the current topics in structural biology and biophysics, which have become central to much of biological research.

细胞质动力蛋白是一种运动蛋白复合物，在许多过程中至关重要，包括有丝分裂期间的染色体分离、囊泡货物的运动、细胞内细胞器的定位以及卵母细胞发育中细胞极性的建立。该项目的重点是表征果蝇动力蛋白轻链 LC8 在动力蛋白组装和货物识别中的作用。 LC8 高度保守，果蝇和人类之间的序列一致性为 94%。动力蛋白中间链IC74介导重链和轻链之间的相互作用，LC8和IC74的组装可能是形成功能复合物的关键步骤。为了阐明 LC8/IC74 识别的基础，将使用 NMR 光谱确定复合物的结构。吞咽蛋白 Swa（货物的一个例子）是一种 RNA 结合蛋白，对于发育中卵母细胞建立细胞极性至关重要。 Swa 与 LC8 的相互作用已在果蝇中得到证实。这种相互作用中结合位点的识别将通过有限的蛋白水解和交联研究来确定。更广泛的影响：LC8 的结构研究及其与 IC74 和 Swa 的相互作用将对理解动力蛋白运动复合物的组装及其与货物的附着做出重大贡献。这项工作将需要许多本科生的实质性贡献。该项目还将涉及结构生物学和生物物理学当前主题的课程的开发，这些主题已成为许多生物学研究的核心。