BRC-BIO: Investigation of the tissue-specific role of the ER membrane protein complex (EMC) in the development and maintenance of a multicellular organism

BRC-BIO：研究 ER 膜蛋白复合物 (EMC) 在多细胞生物体发育和维持中的组织特异性作用

• 批准号: 2313370
• 负责人: Rebecca Delventhal
• 金额: $ 50.26万
• 依托单位: Lake Forest College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2313370&HistoricalAwards=false
• 关键词: BRC; BIO; Investigation; tissue; specific

Cells require many proteins, either embedded in the cellular membrane or moved outside of the cell via small packages, for numerous processes. These proteins can serve a variety of essential functions, such as receptors, tethers, and signals to other cells. These proteins typically are particularly challenging to fold into the correct shape. The endoplasmic reticulum (ER) is the location where this processing occurs with the help of specialized protein complexes. The PI and undergraduate researchers recently found that loss of one component of an ER membrane protein complex, EMC4, in specialized cells of the nervous system called glia, led to a shortened lifespan, locomotor deficits, and other detrimental outcomes in the model organism, the fruit fly. The proposed research will investigate novel questions regarding (1) when during the life cycle is this component required for organismal health, (2) which types of cells in the nervous system require this component to function normally, and (3) what is the role of other members of this ER membrane protein complex. Answering these questions will provide insight into how the ER processes proteins and whether this differs between specialized cell types or life stages of an organism. This project will provide scientific training to Lake Forest College undergraduates through mentored research experiences, improving their access to and preparation for STEM careers. The proposed project will also incorporate a multi-year Course-Based Undergraduate Research Experience (CURE) to significantly increase the number of students that gain valuable research training. The recently discovered ER membrane protein complex (EMC) consists of 8-10 conserved subunits, and is thought to be involved in insertion, folding, and protection from degradation of membrane proteins. However, understanding of EMC function is limited by the fact that most prior work has been conducted in single-cell systems. A multicellular organism is required to investigate the cell type-specific roles of EMC subunits or the impact of EMC function on different life stages. Drosophila represent a powerful model system to answer questions that integrate the molecular and cellular function of EMC with its impact on the organism. The PI and undergraduate researchers recently discovered that glia-specific RNAi knockdown (KD) of the fly homolog of an EMC subunit, EMC4, severely reduces adult survival. Flies lacking glial expression of EMC4 also display a mild developmental delay, but no significant decrease in developmental viability. EMC4 glial KD flies also display severe locomotor defects and increased protein aggregation. This research proposes to investigate (1) the role that glial EMC4 plays in different life stages, specifically during development vs. during maintenance of an adult organism, (2) the cell-specific function of EMC4 in specialized cell types of the nervous system, and (3) the specificity of any of these phenotypes to EMC4 vs. other EMC subunits. This research will advance understanding of the role of EMC in membrane protein biogenesis by taking an integrative approach examining survival, behavior, and development alongside molecular and cellular function.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

细胞需要许多蛋白质，要么嵌入细胞膜中，要么通过小包装在细胞外移动，以进行多种过程。这些蛋白质可以发挥各种必不可少的功能，例如受体，系tethers和信号。这些蛋白质通常特别具有挑战性地折叠成正确的形状。内质网（ER）是该加工借助专用蛋白质复合物进行的位置。 PI和本科研究人员最近发现，在神经系统的专门细胞中，ER膜蛋白复合物的一种成分损失导致寿命缩短，运动型缺陷以及模型有机体中的其他有害结果。拟议的研究将调查有关（1）生命周期时的新问题，当时有机体健康所需的组成部分，（2）神经系统中哪些细胞需要正常起作用，以及（3）该ER膜蛋白蛋白复合物的其他成员的作用是什么。回答这些问题将提供有关ER处理蛋白质的洞察力，以及这在有机体的专业细胞类型或生命阶段之间是否有所不同。该项目将通过指导的研究经验为湖森林学院的本科生提供科学培训，改善其进入和为STEM职业做准备。拟议的项目还将纳入基于多年的课程的本科研究经验（CURE），以显着增加获得宝贵的研究培训的学生人数。 最近发现的ER膜蛋白复合物（EMC）由8-10个保守的亚基组成，被认为参与插入，折叠和保护膜蛋白的降解。但是，对EMC功能的理解受到以下事实的限制：大多数先前的工作是在单细胞系统中进行的。需要多细胞生物来研究EMC亚基的细胞类型特异性作用或EMC功能对不同生命阶段的影响。果蝇代表了一个强大的模型系统，可以回答整合EMC的分子和细胞功能及其对生物体的影响的问题。 PI和本科研究人员最近发现，EMC亚基EMC4的苍蝇同源物的胶质特异性RNAi敲低（KD）严重降低了成人的生存。缺乏EMC4神经胶质表达的苍蝇也显示出轻度的发育延迟，但发育生存能力没有显着降低。 EMC4 Glial KD蝇还显示出严重的运动缺陷和蛋白质聚集的增加。这项研究建议（1）神经胶质EMC4在不同的生命阶段中发挥作用，特别是在发展过程中与维持成人生物体期间的作用，（2）EMC4在神经系统的专用细胞类型中的细胞特异性功能，以及（3）这些表型对EMC4对EMC4与其他EMC子nits的特异性。这项研究将通过采用一种综合方法来研究EMC在膜蛋白生物发生中的作用，并采用一种综合方法，研究生存，行为和发育以及分子和细胞功能。该奖项反映了NSF的法定任务，并被认为是值得通过基金会的知识分子优点和更广泛影响的审查标准来通过评估来支持的。