Clu forms a dynamic new ribonucleoprotein particle directly involved in mitochondrial function

项目摘要：功能齐全的线粒体对细胞至关重要，尤其是 对肌肉、神经元和发育中的胚胎等高能量需求的组织非常重要。AS 这种线粒体功能障碍通常与疾病有关，如神经退行性变、心血管疾病 疾病和糖尿病。线粒体与细胞核协同工作，必须输入数百个细胞核- 编码产物来补充它们自己的小基因组。我们开发了果蝇作为一个模型系统来 研究线粒体，我们的长期目标是了解控制线粒体的分子机制 线粒体在组织动态平衡和发育过程中的功能。我们发现了果蝇蛋白质 无线索(Clu)是支持线粒体功能所必需的。CLU从酵母到人类高度保守， CLU家族成员是核糖核蛋白，它优先结合核编码的mRNAs，目的是 线粒体。CLU在细胞质中形成动态的、大型的线粒体相关颗粒。我们的大妈 这项提案的目标是确定粒子是如何形成的，以及它们在支持 线粒体功能和蛋白质输入。果蝇CLU突变体因受损而患病和不育 线粒体。CLU与位于线粒体外膜的蛋白质结合，包括 负责蛋白质输入的转位酶。CLU还与外膜上的核糖体结合。我们 假设Clu颗粒的形成对调节mRNA的定位和蛋白质的输入是重要的。 为了验证这一假设，我们的第一个目标是定义Clu粒子动力学以及使用 显微镜、遗传学和分子生物学。我们的第二个目标是研究Clu颗粒如何对信使核糖核酸做出贡献 利用显微镜、分子生物学和生物化学进行调控。我们的第三个目标是使用分子生物学， 生物化学和遗传学研究Clu颗粒如何调节线粒体蛋白质水平。《知识》 我们希望从这项关于Clu颗粒监管的提案中获益将是向前迈出的变革性的一步，因为 对参与线粒体蛋白输入的核糖核蛋白以及Clu颗粒知之甚少 代表一种对线粒体功能至关重要的未定义的、新的细胞质颗粒。从以下方面获得的知识 这里提出的研究将极大地促进我们对线粒体定位的理解 对mRNAs进行监管、翻译和进口，这是一个基本的、基本的过程，对 线粒体在所有细胞中都有功能。

PROJECT SUMMARY: Fully functional mitochondria are critically important for cells, and particularly important for tissues with high-energy demands such as muscle, neurons, and the developing embryo. As such, mitochondrial dysfunction is often associated with disease, such as neurodegeneration, cardiovascular disease and diabetes. Mitochondria work in concert with the nucleus and must import hundreds of nucleus- encoded products to supplement their own small genome. We have developed Drosophila as a model system to study mitochondria and our long-term goal is to understand the molecular mechanisms that control mitochondrial function during tissue homeostasis and development. We have found the Drosophila protein Clueless (Clu) is required to support mitochondrial function. Clu is highly conserved from yeast to human and Clu family members are ribonucleoproteins that preferentially bind nucleus-encoded mRNAs destined for mitochondria. Clu forms dynamic, large, mitochondria-associated particles in the cytoplasm. Our broad objectives for this proposal are to determine how particles form and what role they play in supporting mitochondrial function and protein import. Drosophila clu mutants are sick and sterile with damaged mitochondria. Clu associates with proteins located in the mitochondrial outer membrane, including the translocase responsible for protein import. Clu also associates with the ribosome at the outer membrane. We hypothesize that Clu particle formation is important for regulating mRNA localization and protein import. To test this hypothesis, our first Aim defines Clu particle dynamics and the signals that regulates it using microscopy, genetics and molecular biology. Our second Aim examines how Clu particles contribute to mRNA regulation using microscopy, molecular biology and biochemistry. Our third Aim uses molecular biology, biochemistry and genetics to examine how Clu particles regulate mitochondrial protein levels. The knowledge we expect to gain from this proposal on Clu particle regulation will be a transformative step forward because so little is known about the ribonucleoproteins involved in mitochondrial protein import, and Clu particles represent an undefined, novel cytoplasmic particle critical for mitochondrial function. Knowledge gained from the research proposed here will significantly advance our understanding of how mitochondria-localized mRNAs are regulated, translated and imported, which is a basic, fundamental process important for mitochondrial function in all cells.