Structure and Mechanism of a Prion-remodeling Factor

朊病毒重塑因子的结构和机制

• 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
• 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' 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

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）：prions是非常规，高度感染的药物，这些药物完全由采用异常构象的蛋白质组成。在哺乳动物中，Prion介导的感染导致了几种毁灭性和不变的致命神经退行性疾病，统称为可传播的海绵状脑病。王室疾病的标志是淀粉样蛋白的存在，淀粉样蛋白也与非普里安疾病的病理有关，从阿尔茨海默氏病和亨廷顿氏病到全身性淀粉样变性。广泛的长期研究目标是揭示分子伴侣在prion复制中的功能作用。酵母提供了一个极好的范式来研究prion复制机制。 [PSI+]是一种酵母菌prion，可以增加无意码密码子的翻译通读。像哺乳动物的prions一样，[PSI+]完全由蛋白质组成，并通过自我复制的淀粉样蛋白构象体形成，其进化保守的翻译终止因子SUP35P（ERF3）形成。 [PSI+]的遗传和维护受HSP104（一种600 kDa，环形成ATP依赖性的，蛋白质重塑的机器）的控制，该机器与HSP70伴侣系统合作，在prion复制和蛋白质分析中。这项研究的目的是提供对HSP104及其细菌同源物CLPB的prion型复制和蛋白质分解活性的详细机械理解。提出了三个具体目的：1）确定Hsp104-Substrate复合物的3D结构，2）研究HSP104和HSP70/HSP40之间的协同相互作用，以及3），以阐明HSP104 Bi-Chaperone系统的蛋白质分解和PRION重复的机理。为了解决我们的研究问题，我们将使用由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组成的多面方法。这些方法的组合提供了一种强大的方法，可以使新的机械洞察力洞悉这种非依赖ATP依赖性分子机器家族的结构 - 功能关系，以便可以将这些信息用于工程师与生物技术和纳米 - 纳米 - 氨基氨酸中潜在应用的新型生物学活动的新型纳米机器。