Regulation of Proteasome Capacity

项目摘要/摘要 (来自原始应用)协调调节蛋白质合成、运输、翻译后 修饰和降解是细胞功能的基本要求。蛋白质在体内的靶向降解 真核细胞在很大程度上是由蛋白酶体进行的。蛋白酶体允许蛋白质的动态调节 稳定，并通过去除有缺陷的蛋白质来加强蛋白质质量控制。重要的是，对蛋白质的需求 个别细胞内蛋白酶体的降解可因内源信号或外源信号而有所不同 压力源。因此，特定条件下调节蛋白酶体的能力维持充足，但要避免 蛋白质的过度降解是确保正常发育和生理所必需的。未能做到 适当调节蛋白酶体与疾病有牵连。例如，蛋白酶体能力不足是 与神经退行性疾病有关，而蛋白酶体水平和活性通常在 癌细胞。我们不了解细胞如何动态地维持适当的蛋白酶体能力来 满足不断变化的蜂窝需求。关键的是，我们不理解这些疾病的失败或过度活跃 机制导致了疾病。在这里，我提出了一种在线虫中发现 控制蛋白酶体能力的因素，了解它们在分子水平上的工作方式，揭示它们的作用 在正常发育中，并开始确定它们可能如何减轻或促进疾病。我们将(1)使用 简单的基于GFP的记者分析执行大规模的基因筛查，将全面识别 蛋白酶体容量调节因子和(2)我们通过 这些屏幕。(3)我们将揭示特定蛋白酶体亚基的改变如何差异化地改变蛋白质 降解能力、发育和蛋白酶体抑制剂抗性。这项工作将提高我们的 了解在整个动物中调节蛋白酶体能力的基本细胞机制 背景。从长远来看，这些见解将形成药物干预的概念基础， 通过调节细胞蛋白质降解能力来改善人类健康。

Project Summary/Abstract (From the original application) Coordinated regulation of protein synthesis, trafficking, post-translational modification and degradation is a basic requirement for cellular function. Targeted protein degradation in eukaryotic cells is largely carried out by the proteasome. The proteasome allows dynamic regulation of protein stability and enforces protein quality control by removing defective proteins. Importantly, demand for protein degradation by the proteasome within individual cells can vary due to endogenous signals or exogenous stressors. Thus, condition-specific regulation of proteasome capacity to maintain adequate, but avoid excessive, protein degradation is needed to ensure normal development and physiology. Failure to appropriately regulate the proteasome is implicated in disease. For example, deficient proteasome capacity is associated with neurodegenerative conditions, whereas proteasome levels and activity are often elevated in cancerous cells. We do not understand how cells dynamically maintain appropriate proteasome capacity to meet changing cellular needs. Critically, we do not understand how the failure or hyperactivity of these mechanisms contributes to disease. Here I propose a forward genetic approach in C. elegans to discover the factors that control proteasome capacity, understand how they work at the molecular level, uncover their roles in normal development, and begin to identify how they may mitigate or contribute to disease. We will (1) use simple GFP-based reporter assays to perform large-scale genetic screens that will comprehensively identify proteasome capacity regulators and (2) characterize novel proteasome regulators we have discovered through these screens. (3) We will uncover how alterations to specific proteasome subunits differentially alter protein degradation capacity, development, and proteasome inhibitor resistance. This work will improve our understanding of the fundamental cellular mechanisms that regulate proteasome capacity in a whole animal context. In the long-term, these insights will form the conceptual basis for pharmacological interventions that improve human health via modulation of cellular protein degradation capacity.