J-PROTEIN REGULATION OF YEAST PRION PROPAGATION

J 蛋白对酵母朊病毒传播的调节

• 批准号: 8687796
• 负责人: Justin Keith Hines
• 金额: $ 26.82万
• 依托单位: LAFAYETTE COLLEGE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8687796
• 关键词: Alzheimer' s Disease; Amino Acid Sequence; Amino Acids; Amyloid; Amyloid Proteins; Auxilins; Behavior; Binding; Biochemical; Biochemical Phenomena; Biological Models; Biology; Buffers; Complex; Cytosol; Data; Databases; Disease; Elements; Evaluation; Genetic; Goals; Hand; Homologous Gene; Human; Intervention; Knowledge; Laboratories; Life; Literature; Location; Maintenance; Mediating; Methods; Mission; Mitosis; Modeling; Molecular; Molecular Chaperones; National Institute of General Medical Sciences; Neurodegenerative Disorders; Outcome; Parkinson Disease; Pathway interactions; Pattern; Peptide Sequence Determination; Peptides; Prevalence; Prion Diseases; Prions; Process; Proteins; Public Health; Regulation; Research; Role; Saccharomyces cerevisiae; Saccharomycetales; Specificity; Structure; System; Testing; United States National Institutes of Health; Variant; Viral Proteins; Virus; Work; Yeasts; amyloid structure; base; chaperone machinery; daughter cell; genetic manipulation; in vivo; innovation; prion seeds; protein aggregate; protein function; protein misfolding; public health relevance; therapy development; transmission process; yeast genetics; yeast prion

DESCRIPTION (provided by applicant): A critical barrier to advancing the understanding of chaperone function in prion biology is that the fundamental chaperone requirements for most yeast prions remain unidentified. Existing knowledge is disjointed due to a lack of systematic evaluation that controls for variation in yeast strain background and prion structure. The continued existence of this barrier is an important problem because, until it is overcome, an understanding of how protein sequences give rise to amyloids with distinct patterns of chaperone interaction cannot be fully realized. The long-term goal is to utilize the highly tractable budding yeast, S. cerevisiae, to systematically decipher the complex relationships between amyloid-forming yeast prions and molecular chaperone proteins with a goal of better understanding J-protein chaperone function and prion behavior. The objective of this particular application is to determine the functional elements involved in two specific prion-chaperone interactions and to utilize a newly developed genetic system to evaluate chaperone requirements in a tightly controlled eukaryotic model. The central hypothesis is that differences in amyloid structure, arising primarily from amino acid composition, create distinct challenges for prion transmission which are overcome by specific J-protein functions that buffer prions against loss during mitosis. The hypothesis has been formulated on the basis of data produced in the applicant's laboratory. The rationale for the proposed research is that unambiguous determinations of J-protein functional requirements are a necessary step toward understanding the mechanisms of J-protein function in amyloid biology. Using two distinct yeast prions, [URE3] and [SWI+], and the yeast cytosol as model systems, this hypothesis will be tested by pursuing three specific aims: 1) Identify the functional role of the auxilin homolog Swa2 in [URE3] prion propagation; 2) Determine the structural elements responsible for the specificity of the J-protein Ydj1 toward the prion [SWI+]; and 3) Determine whether Asn-content is the primary determinant of secondary J-protein requirements among prion-forming proteins in yeast. Proven yeast genetic manipulations, which have been established as feasible in the applicant's hands, will be the primary methods used to accomplish these aims. The approach is innovative because it represents a substantive departure from the status quo by placing emphasis on the ability to draw distinctions and make comparisons among multiple J-proteins and yeast prions as a way to broadly understand J-protein function. The contribution of the proposed research is expected to be the elucidation of the roles of two distinct J-proteins in prion propagation and the identification of new prion-chaperone requirements. This contribution is significant because it is the first step in a continuum of research which is expected to contribute to the understanding of the biochemical basis of J-protein-amyloid interactions. A molecular understanding of prion-chaperone interactions has the potential to inform the development of interventions for protein misfolding disorders, including the increasing prevalent neurodegenerative disorders Alzheimer's and Parkinson's.

描述（由申请人提供）：推进对朊病毒生物学中伴侣蛋白功能的理解的一个关键障碍是大多数酵母朊病毒的基本伴侣蛋白需求仍然未被确定。现有的知识是脱节的，由于缺乏系统的评价，控制酵母菌株背景和朊病毒结构的变化。这一障碍的持续存在是一个重要的问题，因为，直到它被克服，蛋白质序列如何产生具有不同模式的伴侣相互作用的淀粉样蛋白的理解不能完全实现。长期的目标是利用高度易处理的芽殖酵母，S。酿酒酵母，系统地破译淀粉样蛋白形成酵母朊病毒和分子伴侣蛋白之间的复杂关系，目的是更好地了解J蛋白伴侣蛋白的功能和朊病毒的行为。本申请的目的是确定两个特定朊病毒-伴侣蛋白相互作用中涉及的功能元件，并利用新开发的遗传系统来评估严格控制的真核模型中的伴侣蛋白要求。核心假设是淀粉样蛋白结构的差异，主要由氨基酸组成引起，对蛋白质的合成产生了独特的挑战。 朊病毒传播，通过特异性J蛋白功能克服，缓冲朊病毒在有丝分裂过程中的损失。该假设是根据申请人实验室提供的数据提出的。拟议研究的基本原理是，明确确定J蛋白的功能要求是理解淀粉样蛋白生物学中J蛋白功能机制的必要步骤。使用两种不同的酵母朊病毒[URE 3]和[SWI+]以及酵母胞质溶胶作为模型系统，将通过追求三个特定目标来测试该假设：1）鉴定生长素同源物Swa 2在[URE 3]朊病毒繁殖中的功能作用; 2）确定负责J蛋白Ydj 1对朊病毒[SWI+]的特异性的结构元件;和3）确定Asn含量是否是酵母中朊病毒形成蛋白中次级J蛋白需求的主要决定因素。已确定在申请人手中可行的酵母遗传操作将是用于实现这些目标的主要方法。这种方法是创新的，因为它代表了一个实质性的偏离现状，把重点放在能力，画出的区别，并在多种J-蛋白和酵母朊病毒之间进行比较，作为一种方式来广泛了解J-蛋白的功能。拟议中的研究的贡献，预计将是两个不同的J-蛋白在朊病毒繁殖和识别新的朊病毒伴侣的要求的作用的阐明。这一贡献是重要的，因为它是连续研究的第一步，预计将有助于理解J-蛋白-淀粉样蛋白相互作用的生化基础。朊病毒伴侣相互作用的分子理解有可能为蛋白质错误折叠疾病的干预措施的发展提供信息，包括日益流行的神经退行性疾病阿尔茨海默氏症和帕金森氏症。