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

项目概要/母公司奖摘要 我们的实验室专注于参与化学反应的分子机器的结构和功能研究 大分子的修饰。自然发生的化学修饰的大分子发挥至关重要的作用 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。化学修饰的失调与许多人类疾病有关， 癌症和神经退行性疾病。虽然许多已被描述和特征化，但仍有 大量的化学改性系统知之甚少。我们的长期目标是 阐明各种化学修饰系统中分子机器的结构和功能 并开发新的工具和策略，根据我们的研究， 已经学会了。在接下来的五年里，我们将重点研究两个系统，p97相关的泛素化系统， 和Vault相关的ADP核糖基化系统。 已经鉴定了超过30种人p97突变，这些突变与许多疾病相关。 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 拓扑结构通过辅因子; 2）疾病突变的结构和功能后果; 3） p97及其辅助因子抑制剂的作用机制。我们的努力保证了前所未有的洞察力， p97，细胞蛋白质稳态的中心枢纽。 Vault是真核细胞中最大的核糖核蛋白，具有独特的结构。Vault的功能包括： 与癌症耐药性和先天免疫反应有关。最近，主要穹窿蛋白（MVP）被 被鉴定为可以预测流感疫苗接种应答的9个标记基因之一。十分重要 在这次COVID-19大流行期间的疫苗开发中，对Vault分子机制的深入了解 至关重要。我们将重点关注PARP 4，它是穹窿复合物中催化ADP核糖基化的唯一酶。 阐明PARP 4的功能及其与Vault颗粒的相互作用是理解其分子机制的关键。 Vault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.