Genetic interaction (GI) analysis of the mammalian ERAD system

 描述（由申请人提供）：新合成的蛋白质必须折叠成正确的三维构象，以便在细胞中正常发挥功能;然而，蛋白质折叠是一个复杂的过程。 复杂且容易出错的过程。在细胞或生物体中积累的末端错误折叠蛋白质可能具有毒性作用，因此必须被识别并迅速去除。内质网相关降解（ERAD）是一种对细胞内稳态至关重要的蛋白质质量控制系统，可识别并降解内质网（ER）中的末端错误折叠蛋白质。ERAD缺陷与许多人类疾病有关，包括囊性纤维化和神经退行性疾病;因此，了解该系统的分子细节可能会阐明多种疾病的潜在病理学，并揭示治疗这些疾病的新治疗靶点。以前的研究哺乳动物ERAD已经确定了这个系统的一些组件，并建议ERAD功能作为一个动态网络的物理和功能连接的蛋白质复合物。本研究的长期目标是详细了解后生动物ERAD网络的组织，并了解这种组织结构如何使系统能够监测高度多样化的哺乳动物蛋白质组的折叠状态。该建议的直接目标是使用遗传相互作用（GI）映射来执行ERAD网络拓扑结构的系统级分析。具体目标1的重点是进行全基因组RNAi筛选，以确定参与ERAD底物错位的基因。在具体目标2中，ERAD系统中的GI将通过分层聚类进行测量和组织，以定义基因之间的网络组织和关系。在具体目标3中，ERAD网络的动态将使用功能基因组学和差异GI分析进行研究。总之，这些研究将用于构建哺乳动物ERAD系统的动态网络图。

 DESCRIPTION (provided by applicant): Newly synthesized proteins must fold into the correct 3-dimensional conformation in order to function properly in the cell; however, protein folding is a complex and error-prone process. Terminally misfolded proteins that accumulate in the cell or organism can have toxic effects, and therefore must be recognized and promptly removed. ER-associated degradation (ERAD), a protein quality control system that is essential for cellular homeostasis, identifies and degrades terminally misfolded proteins in the endoplasmic reticulum (ER). Defects in ERAD have been linked to numerous human diseases including cystic fibrosis and neurodegenerative disorders; thus, understanding the molecular details of this system may illuminate the underlying pathologies of multiple diseases and reveal new therapeutic targets for the treatment of these diseases. Previous studies of mammalian ERAD have identified some of the components of this system and suggest that ERAD functions as a dynamic network of physically and functionally connected protein complexes. The long-term goal of this study is to gain detailed insight into the organization of the metazoan ERAD network, and understand how this organizational structure allows the system to monitor the folding status of a highly diverse mammalian proteome. The immediate goal of this proposal is to use genetic interaction (GI) mapping to perform a systems-level analysis of the ERAD network topology. The focus of Specific Aim 1 is to perform a genome-wide RNAi screen to identify genes involved in ERAD substrate dislocation. In Specific Aim 2, GIs in the ERAD system will be measured and organized by hierarchical clustering to define network organization and relationships between genes. In Specific Aim 3, the dynamics of the ERAD network will be studied using functional genomics and differential GI analysis. Together, these studies will be used to build a dynamic network map of the mammalian ERAD system.