Structural and Functional Studies of Molecular Machines Involved in Chemical Modifications of Biomolecules.

来自家长奖的项目摘要/摘要 我们的实验室主要研究与化学有关的分子机器的结构和功能。 大分子的修饰。自然发生的大分子化学修饰起着至关重要的作用 在分子生物学的各个方面发挥作用，从转录和翻译调节到功能调节 各种蛋白质的结合。化学修饰的错误调节与许多人类疾病有关，例如 癌症和神经退行性疾病。尽管已经描述和描述了许多，但仍然有 大量的化学修饰系统知之甚少。我们的长期目标是 阐明参与各种化学修饰系统的分子机器的结构和功能 并开发新的工具和战略，以根据我们所做的调整他们对抗相关疾病的活动 学到了什么。在接下来的五年里，我们将专注于两个系统，与p97相关的泛素化系统。 以及与Vault相关的ADP-核糖化系统。 人类p97基因的30多个突变已被鉴定，它们与许多 神经退行性疾病。然而，分子机制仍然难以捉摸。通过结构生物学 方法和蛋白质工程，我们将解决1)p97如何处理不同的泛素链 2)疾病突变的结构和功能后果；3) P97及其辅因子抑制剂的作用机制。我们的努力为我们提供了前所未有的深入了解 P97，细胞蛋白质动态平衡的中心枢纽。 Vault是真核细胞中最大的核糖核蛋白，具有独特的结构。电子仓库的功能有 与癌症的耐药性和先天免疫反应有关。最近，主要的拱顶蛋白(MVP)是 被确定为能够预测流感疫苗接种反应的9个标记基因之一。鉴于这一重要性 在这次新冠肺炎大流行期间，对疫苗研发的了解，对金库病毒分子机制的深刻理解 是至关重要的。我们将重点关注PARP4，它是保险库复合体中唯一催化ADP-核糖化的酶。 阐明PARP4的功能以及它是如何与Vault颗粒相互作用的是理解分子的关键 电子仓库的功能。

PROJECT SUMMARY / ABSTRACT from the Parent Award Our lab focuses on structural and functional studies of molecular machines involved in chemical modifications of macromolecules. Naturally occurring chemical modifications of macromolecules play essential roles in all aspects of molecular biology, from transcriptional and translational regulation to functional modulation of various proteins. Misregulation of the chemical modifications is involved in many human diseases such as cancers and neurodegenerative diseases. Although many have been described and characterized, there are still significant amounts of chemical modification systems that are poorly understood. Our long term goal is to elucidate the structure and function of molecular machines involved in various chemical modification systems and develop new tools and strategies to modulate their activity against the relevant diseases based on what we have learned. In the next five years, we will be focusing on two systems, the p97 related ubiquitination system and the Vault related ADP-ribosylation system. More than thirty mutations of human p97 have been identified, which are associated with a number of neurodegenerative diseases. The molecular mechanism, however, remains elusive. Through structural biology approaches and protein engineering, we will address 1) how p97 processes ubiquitin chains of different topologies through the cofactors; 2) the structural and functional consequences of disease mutations; 3) the mechanism of inhibitors of p97 and its cofactors. Our efforts promise unprecedented insights into the function of p97, a central hub of cellular protein homeostasis. Vault is the largest ribonucleoprotein in eukaryotic cells with a unique structure. The function of Vault has been linked to drug resistance in cancer and innate immune response. Recently, major vault protein (MVP) was identified as one of the 9 marker genes that can predict influenza vaccination responses. Given the importance of vaccine development during this COVID-19 pandemic, a deep understanding of Vault’s molecular mechanism is critical. We are going to focus on PARP4, the only enzyme in the Vault complex catalyzing ADP-ribosylation. Elucidating the function of PARP4 and how it interacts with Vault particle is the key to understand the molecular function of Vault.