The determinants of aggregation and toxicity of prion-like domains

朊病毒样结构域聚集和毒性的决定因素

基本信息

批准号：
9085335
负责人：
Eric D Ross
金额：
$ 29.05万
依托单位：
COLORADO STATE UNIVERSITY
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 2018-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9085335
关键词：
Address Affect Algorithms Amino Acid Sequence Amino Acids Amyloid Amyotrophic Lateral Sclerosis Asparagine Awareness Base Sequence Bioinformatics Biological Assay Biological Models Cytoplasmic Inclusion Data Degenerative Disorder Dementia Development Disease Drosophila genus EWSR1 gene Epigenetic Process Frontotemporal Dementia Frontotemporal Lobar Degenerations Generations Genetic Glutamine Health Human In Vitro Inclusion Bodies Inclusion Body Myopathy with Early-Onset Paget Disease Knowledge Link Methods Molecular Conformation Mutagenesis Mutation Myopathy Neurodegenerative Disorders Organism Osteitis Deformans Prions Process Proteins Regulatory Element Sample Size TAF15 gene Testing Therapeutic Toxic effect Yeasts age related amyloid formation base candidate identification driving force human disease improved in vitro Assay in vivo interest novel prediction algorithm prion-like protein TDP-43 protein aggregation research study success sup35 yeast prion

项目摘要

DESCRIPTION (provided by applicant): Prions are infectious proteins that result from the structural conversion of proteins into a self-templating conformation. In yeast, a number of glutamine/asparagine-rich proteins have been shown to undergo prion conversions from a soluble form to an insoluble amyloid form. A variety of evidence suggests that these prions may act as epigenetic regulatory elements. Interestingly, numerous human proteins also contain prion-like domains (PrLDs) - domains with compositional similarity to the yeast prion-forming domains. Six of these have recently been linked to various age-related degenerative disorders, heightening the interest in PrLDs. More than 200 other human proteins contain PrLDs, suggesting that aggregation of these proteins may be involved in other diseases. However, despite the importance of these domains in human disease, the sequence basis for their aggregation and toxicity is still poorly understood. Preliminary attempts to define the sequence basis for aggregation of PrLDs recently resulted in the development of PAPA (Prion Aggregation Prediction Algorithm), the first algorithm demonstrated to be able to distinguish between proteins with and without prion-like activity. This proposal builds on these early successes, using a combination of yeast and Drosophila genetics, in vitro experiments, and bioinformatics to rigorously define how the amino acid sequence of PrLDs contributes to aggregation and toxicity. In Specific Aim 1, a newly developed assay will be employed to quantitatively determine the compositional requirements for aggregation and toxicity of PrLDs. In Specific Aim 2, a combination of in vitro assays and Drosophila experiments will be used to explore how the propensity to convert to oligomeric and amyloid species affects toxicity in vivo. Finally, in Specific Aim 3, sophisticated bioinformatics methods will be used to incorporate this information into improved prediction algorithms, allowing for the identification of candidate disease-associated or regulatory PrLDs.

描述（由申请人提供）：prions是由蛋白质转化为自我示意构象的结构转化而产生的传染蛋白。在酵母中，已经显示出许多富含谷氨酰胺/天冬酰胺的蛋白质可以从可溶性形式转化为不溶性淀粉样蛋白形式。各种证据表明，这些王室可能充当表观遗传调节元素。有趣的是，许多人类蛋白还包含与酵母菌形成型prion的域组成相似性的域类域（PRLD）。其中六个最近与各种与年龄相关的退化性疾病有关，从而提高了对PRLD的兴趣。其他200多个人类蛋白质含有PRLD，这表明这些蛋白质的聚集可能与其他疾病有关。然而，尽管这些领域在人类疾病中的重要性，但其汇总和毒性的序列基础仍然很少了解。定义PRLD聚集的序列基础的初步尝试导致PAPA的发展（Prion聚集预测算法），第一种算法证明能够区分具有和没有prion活性的蛋白质。该建议是建立在这些早期成功基础上的，结合了酵母和果蝇遗传学，体外实验和生物信息学，以严格地定义PRLD的氨基酸序列如何有助于聚集和毒性。在特定的目标1中，将采用新开发的测定法来定量确定PRLD聚集和毒性的组成要求。在特定的目标2中，将使用体外测定和果蝇实验的组合来探索转化为寡聚和淀粉样物种的倾向如何影响体内的毒性。最后，在特定的目标3中，将使用复杂的生物信息学方法将这些信息纳入改进的预测算法中，从而识别候选疾病相关或调节性PRLD。