Defining the role of site-specific proteolysis in innate defense signaling

项目总结 人类细胞对病原体和毒素等外来物质做出反应，启动了一种强大的天然防御 在细胞中创造保护性环境并使入侵的病原体丧失能力的反应 外来物质。这种先天防御反应的启动、激活和解决是一个谨慎的过程。 受监管的过程，旨在避免过度激活和欠激活，其中任何一种都可能导致 组织损伤、器官功能障碍和微生物疾病。 细胞用来实现对天然防御信号的严格调控的一个关键策略是改变 蛋白质翻译后修饰(PTMS：磷酸化、泛素化、乙酰化等)。的目标是 在接下来的五年里，我的实验室将探索一种未被充分研究的PTM的作用，即位点特异性蛋白分解， 调节先天防御信号。我在研究抗病毒先天免疫方面有丰富的经验 最近优化了一种健壮的蛋白质N末端标记方法，该方法提供了一种 对细胞中特定部位的蛋白分解的公正观点--这一专业知识将使我能够破译 位点特异性蛋白分解在先天防御机制中的作用。 有了这个Mira奖，我的团队将追求两个具体的研究方向。首先，我们将确定角色 蛋白水解性N-端规则通路在控制参与防御信号的蛋白质的稳定性中的作用。 其次，我们将研究内部蛋白质裂解激活防御反应的方式。对于 建议的研究，我们将采用重点分子生物学研究和系统的组合 生物学方法，如基于活性的蛋白酶图谱和蛋白质N末端标记。这些努力 将产生关于蛋白酶在调节细胞防御中的作用的新知识，并告知 制定策略以改进针对升级的先天防御机制的性能 微生物和环境威胁。

PROJECT SUMMARY Human cells respond to foreign agents such as pathogens and toxins by initiating a strong innate defense response that creates a protective environment in the cells and incapacitates the invading pathogens and foreign substances. The initiation, activation, and resolution of this innate defense response is a carefully regulated process designed to avoid both hyperactivation and underactivation, either of which can lead to tissue damage, organ dysfunction, and microbial diseases. A key strategy that cells use to achieve tight regulatory control of innate defense signaling is the alteration of protein post-translational modifications (PTMs: phosphorylation, ubiquitination, acetylation, etc.). The goal of my lab for the next five years will be to explore the role of an understudied PTM, the site-specific proteolysis, in regulating innate defense signaling. I have extensive experience in studying antiviral innate immune responses, and have recently optimized a robust protein N-terminal labeling approach that provides an unbiased view of site-specific proteolysis in cells — expertise that will enable me to decipher the regulatory functions of site-specific proteolysis in innate defense mechanisms. With this MIRA award, my group will pursue two specific research directions. First, we will determine the role of the proteolytic N-end rule pathway in controlling the stability of proteins involved in defense signaling. Second, we will investigate the ways in which internal protein cleavages activate defense responses. For the proposed studies, we will employ a combination of focused molecular biology investigations and systems biology approaches, such as activity-based protease profiling and protein N-terminal labeling. These efforts will generate new knowledge about the role of proteases in the regulation of cellular defenses and inform the development of strategies to improve the performance of innate defense mechanisms against escalating microbial and environmental threats.