Functions of the Mitochondrial Proteome: An Integrated Multi-Species Approach

描述（由申请人提供）：线粒体是动态和复杂的细胞器，在生物学的各个方面发挥核心作用，包括能量产生、中间代谢和细胞凋亡。这些广泛的细胞功能也使线粒体成为人类健康的核心参与者。线粒体功能障碍与多种疾病相关，包括癌症、2型糖尿病和大多数神经退行性疾病。由于这些广泛的关键活动，许多努力都集中在识别和表征线粒体蛋白质组，迄今为止在哺乳动物中鉴定了1,000多种蛋白质。然而，值得注意的是，大约四分之一的这些蛋白质仍然基本上没有特征。这些包括许多在真核生物中高度保守的蛋白质，这强烈表明它们具有重要的功能。我们先前对两种未表征的线粒体保守蛋白（MCP）的研究支持了这一提议，揭示了这些蛋白在线粒体功能的关键方面的新作用，并且在一种情况下，提供了与遗传形式的癌症的直接联系。我们建议继续这些线粒体蛋白质组的功能研究，在酵母，果蝇，哺乳动物细胞和小鼠中使用遗传和生化方法。本提案中的三个具体目标分别集中在线粒体功能的不同关键方面：（1）确定BRP 44蛋白家族在碳水化合物代谢中的作用，（2）确定C6 orf 57蛋白家族在维持能量稳态中的作用，以及（3）确定HIG蛋白在线粒体III 2 IV 2呼吸超复合物组装中的作用。我们将在酵母中进行遗传和生物化学研究，重点是确定每个MCP的机制功能。这些研究将通过果蝇的遗传和代谢研究进行扩展，与我们的酵母工作密切协调，以确定每个MCP在完整发育动物中的生物和生理活性。在哺乳动物细胞中的功能研究将被用来测试特定的MCP活动已通过进化保守的假设。最后，将产生选定的小鼠模型，以阐明蛋白质在哺乳动物生理学中的功能。我们在这项研究中的目标将是提供对每个MCP的生化和细胞功能的新理解，确定它们如何促进正常的线粒体活性，并尽可能将MCP功能与人类健康和疾病联系起来。

DESCRIPTION (provided by applicant): Mitochondria are dynamic and complex organelles that play a central role in all aspects of biology, including energy production, intermediary metabolism, and apoptosis. These broad cellular functions also place mitochondria as a central player in human health. Mitochondrial dysfunction is associated with a wide range of diseases, including cancer, type 2 diabetes, and most neurodegenerative disorders. As a result of these wide-ranging critical activities, many efforts have focused on identifying and characterizing the mitochondrial proteome, with over 1,000 proteins identified to date in mammals. Remarkably, however, roughly one-quarter of these proteins remain essentially uncharacterized. These include many proteins that are highly conserved throughout eukarya, a strong indication that they perform a fundamentally important function. Our prior studies of two uncharacterized mitochondrial conserved proteins (MCPs) support this proposal, revealing new roles for these proteins in critical aspects of mitochondrial function and, in one case, providing a direct link to an inherited form of cancer. We propose to continue these functional studies of the mitochondrial proteome using genetic and biochemical approaches in yeast, Drosophila, mammalian cells, and mice. The three specific aims in this proposal each focus on a different critical aspect of mitochondrial function: (1) To determine the role of the BRP44 protein family in carbohydrate metabolism, (2) To determine the role of C6orf57 protein family in maintaining energy homeostasis, and (3) To determine the role of HIG proteins in mitochondrial III2IV2 respiratory super complex assembly. We will undertake genetic and biochemical studies in yeast focused on defining the mechanistic function of each MCP. These studies will be extended through genetic and metabolic studies in Drosophila, in close coordination with our yeast work, to determine the biological and physiological activity of each MCP in the context of an intact developing animal. Functional studies in mammalian cells will be used to test the hypothesis that specific MCP activities have been conserved through evolution. Finally, selected mouse models will be generated to enable elucidation of protein function in mammalian physiology. Our goal in this research will be to provide a new understanding of the biochemical and cellular function of each MCP, determine how they contribute to normal mitochondrial activity, and, whenever possible, link MCP function to human health and disease.