Structure and Mechanism of a Prion-remodeling Factor

朊病毒重塑因子的结构和机制

基本信息

批准号：
8531529
负责人：
Francis T.F. Tsai
金额：
$ 31.69万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-01 至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8531529
关键词：
Address Adopted Alzheimer&apos s Disease Amyloid Amyloidosis Animal Model Binding Biochemical Biological Biology Biotechnology Bovine Spongiform Encephalopathy Cells Chemicals Complex Disease Engineering Family Genetic Grant Health Homeostasis Homologous Gene Human Huntington Disease Hybrids In Vitro Infection Infectious Agent Longevity Maintenance Mammals Mediating Membrane Methods Molecular Molecular Chaperones Molecular Conformation Molecular Machines Monitor Mutagenesis Neurodegenerative Disorders Nonsense Codon Pathology Peptide Hydrolases Peptides Prion Diseases Prions Property Protein Family Protein Structure Initiative Proteins Proteome Proteomics Reading Recovery of Function Research Rewards Risk Role Sorting - Cell Movement Stress Structure Structure-Activity Relationship Substrate Interaction System Tail Techniques Translations Work Yeasts base biological adaptation to stress conformer in vivo insight nanomachine nanomedicine non-prion novel polypeptide prevent protein aggregate protein aggregation protein folding protein misfolding public health relevance structural biology termination factor three dimensional structure yeast genetics yeast prion

项目摘要

DESCRIPTION (provided by applicant): Prions are unconventional, highly infectious agents, which are composed entirely of a protein that adopts an abnormal conformation. In mammals, prion-mediated infections are responsible for several devastating and invariably fatal neurodegenerative diseases, collectively known as transmissible spongiform encephalopathies. A hallmark of prion diseases is the presence of amyloids, which are also associated with the pathology of non-prion diseases, ranging from Alzheimer's and Huntington's disease to systemic amyloidosis. The broad and long-term research objective is to uncover the functional role of molecular chaperones in prion replication. Yeast provides an excellent paradigm to investigate the mechanism of prion replication. [PSI+] is a yeast prion that increases translational read-through of nonsense codons. Like mammalian prions, [PSI+] consist entirely of protein and is formed by self-replicating amyloid conformers of the evolutionary conserved translation termination factor Sup35p (eRF3). The inheritance and maintenance of [PSI+] are governed by Hsp104, a 600-kDa, ring-forming ATP-dependent, protein-remodeling machine, which cooperates with the Hsp70 chaperone system in prion replication and protein disaggregation. The objective of this research is to provide a detailed mechanistic understanding of the prion-remodeling and protein disaggregating activities of Hsp104 and its bacterial homolog ClpB. Three specific aims are proposed: 1) to determine the 3D structure of an Hsp104-substrate complex, 2) to investigate the synergistic interaction between Hsp104 and Hsp70/Hsp40, and 3) to elucidate the mechanism of protein disaggregation and prion replication by the Hsp104 bi-chaperone system. To address our research questions, we will use a multi-facet approach consisting of hybrid structural biology methods, proteomic and chemical biology techniques, and yeast genetics. The combination of these methods provides a powerful approach to yield new mechanistic insight into the structure-function relationship of this remarkable family of ATP-dependent molecular machines in order that this information might be exploited to engineer new nano-machines with novel biological activities with potential applications in biotechnology and nano-medicine.

描述（由申请人提供）：朊病毒是非常规的、高度传染性的病原体，其完全由采用异常构象的蛋白质组成。在哺乳动物中，朊病毒介导的感染导致几种毁灭性且致命的神经退行性疾病，统称为传染性海绵状脑病。朊病毒疾病的一个标志是淀粉样蛋白的存在，淀粉样蛋白也与非朊病毒疾病的病理学相关，从阿尔茨海默病、亨廷顿病到系统性淀粉样变性。广泛而长期的研究目标是揭示分子伴侣在朊病毒复制中的功能作用。酵母为研究朊病毒复制机制提供了一个极好的范例。 [PSI+] 是一种酵母朊病毒，可增加无义密码子的翻译通读。与哺乳动物朊病毒一样，[PSI+] 完全由蛋白质组成，由进化保守的翻译终止因子 Sup35p (eRF3) 的自我复制淀粉样蛋白构象异构体形成。 [PSI+] 的遗传和维持由 Hsp104 控制，Hsp104 是一种 600 kDa、成环 ATP 依赖性蛋白质重塑机器，它与 Hsp70 伴侣系统合作进行朊病毒复制和蛋白质解聚。本研究的目的是提供对 Hsp104 及其细菌同源物 ClpB 的朊病毒重塑和蛋白质解聚活性的详细机制了解。提出了三个具体目标：1) 确定 Hsp104-底物复合物的 3D 结构，2) 研究 Hsp104 和 Hsp70/Hsp40 之间的协同相互作用，3) 阐明 Hsp104 蛋白质解聚和朊病毒复制的机制双伴侣系统。为了解决我们的研究问题，我们将采用多方面的方法，包括混合结构生物学方法、蛋白质组学和化学生物学技术以及酵母遗传学。这些方法的结合提供了一种强大的方法，可以对这一非凡的 ATP 依赖性分子机器家族的结构-功能关系产生新的机制洞察，以便利用这些信息来设计具有潜在生物活性的新型纳米机器。生物技术和纳米医学中的应用。