Visualizing the Mechanisms of Protein Quality Control

蛋白质质量控​​制机制的可视化

• 批准号: 10574767
• 负责人: Peter Shen
• 金额: $ 7.31万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10574767
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; Address; Autophagocytosis; Binding; Cell Cycle Progression; Cell physiology; Cells; Clinical; Complex; Cryoelectron Microscopy; Cues; Degenerative Disorder; Enzymes; Equilibrium; Functional disorder; Gene Expression; Goals; Human; Image; Inclusion Body Myopathy with Early-Onset Paget Disease; Life; Malignant Neoplasms; Membrane Fusion; Methods; Molecular; Molecular Conformation; Molecular Machines; Mutation; Parents; Pathway interactions; Protein Biosynthesis; Proteins; Proteomics; Quality Control; Research; Role; Therapeutic; Work; cancer therapy; cofactor; familial amyotrophic lateral sclerosis; inhibitor; programs; proteostasis; tool; trafficking; tumor; valosin-containing protein

Project Summary / Abstract Optimal cellular function requires balanced networks that maintain protein synthesis, folding, trafficking, remodeling and degradation. The Cdc48/p97/VCP AAA ATPase is an essential and abundant molecular machine that helps maintain this balance across eukaryotic life and is therefore a critical control point for proteostasis. Cdc48 is best characterized for its role in targeting ubiquitylated proteins for proteasomal degradation, but the enzyme also functions in a wide range of other essential pathways, including cell cycle progression, autophagy, membrane fusion, and gene expression. Mutations in human Cdc48 are causative of a multisystem proteopathy that clinically manifests as a combination of Inclusion Body Myopathy, Paget's Disease of Bone, Frontotemporal Dementia (collectively known as IBMPFD), and familial Amyotrophic Lateral Sclerosis (fALS). Moreover, Cdc48 expression is elevated in several tumors and its inhibitors are an emerging class of therapeutics for cancer treatment. Despite these critical roles, surprisingly little is known about the molecular mechanisms that allow Cdc48 to perform its myriad cellular functions. More than thirty adaptors and cofactors are known to interact with Cdc48, but how cells organize these binding partners into functional complexes is also unknown. My research program aims to define the mechanisms underlying Cdc48 functions by using an integrative approach of endogenous purification, proteomics, cryo-EM imaging, and computational processing methods. Our approach will be used to visualize and characterize Cdc48 assemblies in an array of its native compositional and conformational states. During the project period, we propose to achieve two major goals: first, we aim to determine the molecular cues that functionally separate Cdc48 across multiple cellular pathways; second, we seek to resolve how Cdc48 converts energy from ATP hydrolysis into a pulling force that remodels and unfolds its protein substrates. Addressing these questions is essential to understand how Cdc48 drives a wide range of cellular processes and how its dysfunction and misregulation contribute to degenerative disease and cancer. The tools we develop to accomplish these goals will likely be broadly applicable in defining the structural landscapes of other challenging molecular machines.

项目摘要/摘要