Molecular Dissection of Mitochondria-Organelle Interactions

项目总结/摘要 线粒体是一种多功能的细胞器，在代谢、蛋白质稳态、钙离子代谢和细胞内环境中起着重要作用。 信号、免疫和细胞死亡。这些线粒体功能的扰动是一个潜在的特征， 几种常见的疾病和生理状况，如癌症、神经退行性疾病和 衰老线粒体与其他细胞器的相互作用在线粒体的调节中起着关键作用。 功能协调发展的然而，这些细胞器-细胞器相互作用仍然知之甚少，部分原因是缺乏 能够分离和表征与不同细胞器相关的线粒体的方法。 这也阻碍了对疾病中的细胞器-细胞器相互作用及其对疾病的贡献的研究。 发病机理该提案的目标是克服这一技术挑战，以研究 在细胞系和细胞模型中通过细胞器相互作用的线粒体调节机制， 线粒体疾病在这里，我建议开发一种创新的生化工作流程， 基于线粒体的细胞位置和细胞器的差异标记和快速分离 交互.使用这种方法，我们将首先表征蛋白质组和代谢物组成， 线粒体与四个细胞器和质膜非常接近。接下来，我们将调查 特定的代谢物和蛋白质修饰是否调节细胞器接触时的线粒体功能 网站.我们还将确定线粒体-细胞器的相互作用是如何改变的细胞模型， 线粒体疾病以及这些相互作用是否可以靶向治疗增益。这些实验 这将是第一次对健康细胞中的细胞器相互作用进行全面的生化分析， 线粒体功能障碍的细胞。我们的研究结果将导致识别信号，源自不同的 细胞结构来调节线粒体功能，并将阐明其机制。此外，本发明还提供了一种方法， 在线粒体功能障碍的细胞中，对线粒体-细胞器相互作用的研究有可能 改变我们对线粒体疾病机制的理解，这将导致新的治疗方法 接近。所开发的方法可以很容易地应用于研究其他细胞器间的相互作用， 以前所未有的生化细节揭示它们的介质和机制。

PROJECT SUMMARY/ ABSTRACT Mitochondria are multi-functional organelles that play central roles in metabolism, protein homeostasis, calcium signaling, immunity, and cell death. Perturbation of these mitochondrial functions is an underlying feature of several common diseases and physiological conditions such as cancer, neurodegenerative diseases and aging. Interaction of mitochondria with other organelles plays a pivotal role in regulation of mitochondrial functions. However these mitochondria-organelle interactions remain poorly understood, partly due to lack of methods that enable isolation and characterization of mitochondria that associate with different organelles. This also hinders investigation of mitochondria-organelle interactions in diseases and their contribution to disease pathogenesis. The goal of this proposal is to overcome this technical challenge in order to investigate the mechanisms of mitochondria regulation through organelle interactions in cell lines and cellular models of mitochondrial diseases. Here, I propose the development of an innovative biochemical workflow that will allow differential tagging and rapid isolation of mitochondria based on their cellular location and organellar interactions. Using this method, we will first characterize the proteome and metabolite composition of mitochondria that are in close proximity to four organelles and the plasma membrane. Next, we will investigate whether specific metabolites and protein modifications regulate mitochondrial functions at organelle contact sites. We will also determine how mitochondria- organelle interactions are altered in cellular models of mitochondrial disease and whether these interactions can be targeted for therapeutic gain. These experiments will be the first comprehensive biochemical analysis of mitochondria-organelle interactions in healthy cells and cells with mitochondrial dysfunction. Our results will lead to identification of signals that originate from different cellular structures to regulate mitochondrial functions and will elucidate their mechanisms. In addition, investigation of mitochondria-organelle interactions in cells with mitochondrial dysfunction has the potential to transform our understanding of mechanisms of mitochondrial diseases, which will lead to novel treatment approaches. The methods developed can easily be applied to investigate other inter-organellar interactions to reveal their mediators and mechanisms at unprecedented biochemical detail.