Regulation of Proteasome Capacity

项目摘要 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。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。蛋白酶体允许动态的 调节蛋白质的稳定性，并通过去除缺陷蛋白质来实施蛋白质质量控制 proteins.重要的是，个体内蛋白酶体对蛋白质降解的需求 细胞可以由于内源性信号或外源性应激源而变化。因此，特定条件 调节蛋白酶体的能力，以保持足够的，但避免过量的蛋白质 需要降解以确保正常发育和生理。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与神经退行性疾病相关。 通常在癌细胞中升高。我们不知道细胞是如何动态维持 适当的蛋白酶体能力，以满足不断变化的细胞需求。关键是，我们不 了解这些机制的失效或过度活跃如何导致疾病。这里我 提出了一种C.来发现那些控制 蛋白酶体的能力，了解他们如何在分子水平上工作，揭示他们的作用， 正常发育，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的报告基因检测来进行大规模的遗传筛查， 全面鉴定蛋白酶体容量调节剂和（2）表征新的 我们通过这些筛选发现的蛋白酶体调节因子。(3)我们将揭开 特异性蛋白酶体亚基的改变差异性地改变蛋白质降解能力， 发育和蛋白酶体抑制剂抗性。这项工作将增进我们对 调节整个动物体内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的概念基础。 通过调节细胞蛋白质降解改善人类健康的干预措施 容量

PROJECT SUMMARY 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.