Regulation of Proteasome Capacity

项目总结/摘要 (From原申请）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。靶向蛋白降解 真核细胞中的蛋白酶主要是由蛋白酶体进行的。蛋白酶体允许蛋白质的动态调节 稳定性，并通过去除缺陷蛋白质来实施蛋白质质量控制。重要的是，对蛋白质的需求 单个细胞内蛋白酶体的降解可由于内源性信号或外源性信号而变化。 压力源因此，条件特异性调节蛋白酶体的能力，以保持足够的，但避免 需要过度的蛋白质降解来确保正常的发育和生理。未能 适当调节蛋白酶体与疾病有关。例如，蛋白酶体能力不足， 与神经退行性疾病相关，而蛋白酶体水平和活性通常在神经退行性疾病中升高。 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， 满足不断变化的蜂窝需求。关键的是，我们不明白这些失败或过度活跃是如何造成的。 导致疾病的机制。在这里，我提出了一个前向遗传方法在C。优雅的人去发现 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， 并开始确定它们如何减轻或促进疾病。我们将（1）使用 简单的基于GFP的报告基因检测，以进行大规模的遗传筛选， 蛋白酶体容量调节剂和（2）表征我们通过以下发现的新型蛋白酶体调节剂： 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 了解调节整个动物蛋白酶体能力的基本细胞机制 上下文从长远来看，这些见解将形成药理学干预的概念基础， 通过调节细胞蛋白质降解能力改善人类健康。

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.