Uncovering novel mechanisms of the CELF/Bruno protein ETR-1 in apoptosis

项目总结/摘要： 细胞程序性死亡或凋亡是一种自然的发育过程。凋亡的失败可导致 发育异常，癌症（不受控制的生长）和自身免疫性疾病，而过度 细胞凋亡导致神经退行性疾病，如阿尔茨海默氏症和帕金森氏症。新玩家 细胞凋亡的各个阶段（启动、执行和吞噬）继续被鉴定，表明我们的研究结果表明， 仍然缺乏对细胞凋亡发生和调节的机制的理解。C. elegans是一个 理想的遗传系统，以解决凋亡的研究问题，因为透明的动物允许 真实的时间，在体内，观察生殖系中的细胞凋亡（生理或应激诱导的生殖系细胞凋亡）。 我们最近在C.优雅的属于高度 保守的CELF/Bruno蛋白家族ETR-1在生殖细胞凋亡中起作用。初步调查结果 显示ETR-1的缺失导致生殖系凋亡细胞的积累，并且 凋亡吞噬途径是ETR-1的潜在mRNA靶点。拟议的总体目标 本研究旨在阐明ETR-1在生殖细胞凋亡过程中的作用及其作用机制 在凋亡过程中。指导所提出的工作的中心假设是ETR-1在RNA结合中起作用。 依赖的方式在早期阶段的生理生殖细胞凋亡，以调节执行， 在吞噬的后期这项研究的基本原理是，成功完成将有助于 我们对细胞凋亡在多个层面上的基本理解。为了验证中心假设，我们设计了 两个具体目标。具体目标1将侧重于确定ETR-1在生理或应激中的作用- 通过检测ETR-1耗竭后细胞凋亡的动力学和组织特异性 性腺中ETR-1的需求与细胞凋亡有关。具体目标2将侧重于了解 生殖细胞凋亡过程中ETR-1的RNA识别基序（RRM）结构域的功能。拟议工作 是创新的，因为我们将在体内阐明一种新的凋亡参与者在完整的多细胞 生物体，并确定以前未表征的RNA结合蛋白的mRNA靶点。完成 这些目标将提供对细胞凋亡过程以及细胞凋亡是如何 适当控制。这项提案的另一个重要的长期目标是培养本科生 对生物医学研究领域感兴趣。这一建议将促进以下方面的实质性参与： 在霍华德大学，一个主要的HBCU， （历史上的黑人大学）。这项研究具有很强的

PROJECT SUMMARY/ABSTRACT: Programmed cell death or apoptosis is a natural developmental process. Failure of apoptosis can lead to developmental abnormalities, cancer (uncontrolled growth) and autoimmune diseases, while excessive apoptosis contributes to neurodegenerative diseases like Alzheimer's and Parkinson's. New players in the various stages of apoptosis (initiation, execution, and engulfment) continue to be identified, indicating that our understanding of the mechanism by which apoptosis occurs and is regulated is still lacking. C. elegans is an ideally suited genetic system to address apoptotic research questions because the transparent animals allows real time, in vivo, observation of apoptosis in the germline (physiological or stress-induced germline apoptosis). We recently identified a well-conserved RNA-binding protein (RBP) in C. elegans belonging to the highly conserved CELF/Bruno protein family, ETR-1, as playing a role in germline apoptosis. Preliminary findings show that depletion of ETR-1 results in accumulation of germline apoptotic cells and that a member of the apoptotic engulfment pathway is a potential mRNA target of ETR-1. The overall objective of this proposed study is to elucidate ETR-1's function during germline apoptosis, and the mechanism by which ETR-1 acts during apoptosis. The central hypothesis guiding the proposed work is that ETR-1 functions in a RNA-binding dependent manner during the early stages of physiological germline apoptosis to regulate execution and also in the later stage of engulfment. The rationale for this research is that successful completion will contribute to our fundamental understanding of apoptosis at multiple levels. To test the central hypothesis, we have devised two specific aims. Specific Aim 1 will focus on determining the role of ETR-1 in either physiological or stress- induced apoptosis through examining the kinetics of apoptosis upon ETR-1 depletion and the tissue specific requirement of ETR-1 in the gonad as related to apoptosis. Specific Aim 2 will focus on understanding the function of the RNA Recognition Motif (RRM) domains of ETR-1 during germline apoptosis. The proposed work is innovative because we will be elucidating in vivo the role for a novel apoptotic player in an intact multicellular organism, and identifying mRNA targets for a previously uncharacterized RNA-binding protein. Completion of these aims will provide a more thorough understanding of the process of apoptosis, and how apoptosis is properly controlled. Another important long-term goal of this proposal is to foster undergraduate student interest in the area of biomedical research. This proposal will facilitate substantial participation of underrepresented minority students in their early career years at Howard University, a major HBCU (Historically Black Colleges and Universities). This research has strong