Amyloids, aggregates and Nucleolar activity in Xenopus development

爪蟾发育中的淀粉样蛋白、聚集体和核仁活性

基本信息

批准号：
10170380
负责人：
DANIEL L. WEEKS
金额：
$ 30.4万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-05 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10170380
关键词：
Address Alzheimer like pathology Alzheimer&apos s Disease Amyloid Biochemical Biological Cataract Cell Nucleolus Cell Nucleus Cell division Cell physiology Cells Characteristics Cytosol DNA Damage DNA-Directed RNA Polymerase Development Disease Ensure Environment Equilibrium Experimental Models FBXW7 gene Foundations Fractionation Gel Gene Expression Regulation Genetic Processes Genetic Transcription Genetic Translation Homeostasis Hormones Huntington Disease Immunohistochemistry Liquid substance Maintenance Mediating Memory Methods Microscopy Modeling NPM1 gene Nature Normal Cell Nuclear Oocytes Oogenesis Organelles Pathogenesis Pathology Phase Physiological Play Post-Translational Protein Processing Property Proteins Proteomics RNA RNA Helicase RNA Processing Rana Recording of previous events Regulation Ribosomal RNA Ribosomes Role Solid Stress Structure System Testing Xenopus laevis Xenopus oocyte amyloid formation base biological adaptation to stress design fibrillarin genetic information helicase in vivo long term memory nucleophosmin particle protein aggregation protein structure response targeted treatment transcriptome sequencing transcriptomics xenopus development

项目摘要

Project Summary: Many cellular processes critical for normal development have devastating consequences when they are not controlled. Not too long ago, protein aggregation was largely equated with pathogenesis. More recently we know that proteins aggregate to mediate stress responsiveness, cell division, long-term memory and to self-assemble into critical non-membrane bound particles in the nucleus and cytosol. If fact, at least 5% of known proteins have the composition that would allow them to form aggregates. Lack of proper cellular control of protein aggregation leads to amyloid formation, which unregulated, is part of the pathogenesis that accompanies devastating diseases like Alzheimer’s and Huntington’s disease and common debilitating conditions like cataracts. The physical nature of aggregates may be that of phase separated liquid droplets, gels, disordered aggregates and the ordered aggregates characteristic of amyloid. Aggregates serve to greatly increase the local concentration of the aggregating protein, and are one part of an equilibrium between different structural states. Consequently, the ability and rate of proteins joining or leaving an aggregate is an important feature that can be regulated, and if compromised leads to aggregate based pathology. There is a gap in understanding which proteins participate in aggregate formation and how aggregation is regulated in vivo. We propose that an ideal system to begin to bridge the gap would be the oocytes of the frog Xenopus laevis. We and others have shown that aggregates accumulate during oogenesis in the cytosol and in nucleus. The nuclear particles responsible for RNA transcription and processing by all three RNA polymerases self-assemble into aggregates, generally with RNA as a co-aggregate. The focus of this proposal is on nucleolus, a prominent non-membrane bound particle responsible for synthesis and processing of ribosomal RNA, the manufacturing of ribosomes, helping to mediate cellular responses to stress and DNA damage, and appropriately responding to cellular need for new translational capacity. We use microscopy, immunohistochemistry, biochemical fractionation, protein depletion, proteomic and transcriptomic analysis to understand the accumulation, function and stability of the aggregates that establish nucleolar form and function. We take advantage of the unique properties of Xenopus oocyte nuclei, including size, abundant protein and RNA content and ease of manipulation to perform our studies. The findings will impact our understanding of how cells maintain physiologically normal equilibrium between native and aggregating forms of proteins.

项目概要：许多对正常发育至关重要的细胞过程如果被破坏，就会产生毁灭性的后果。不久前，蛋白质聚集在很大程度上与发病机制等同。最近我们知道蛋白质聚集可以调节应激反应、细胞分裂、长期记忆并在细胞核和细胞质中自组装成关键的非膜结合颗粒。事实上，至少 5% 的已知蛋白质具有允许它们形成聚集体的成分。缺乏对蛋白质聚集的适当细胞控制会导致淀粉样蛋白的形成，而淀粉样蛋白的形成不受调节，阿尔茨海默氏症和亨廷顿舞蹈症等毁灭性疾病的部分发病机制疾病和常见的使人衰弱的病症，如白内障，聚集体的物理性质可能是。相分离的液滴、凝胶、无序聚集体和有序聚集体淀粉样蛋白的聚集特性可以大大增加淀粉样蛋白的局部浓度。聚集蛋白质，并且是不同结构状态之间平衡的一部分。经过测试，蛋白质加入或离开聚集体的能力和速率是一个重要特征这是可以调节的，如果受到损害，就会导致基于聚合的病理学上存在差距。了解哪些蛋白质参与聚集体形成以及聚集体是如何调节的我们认为青蛙的卵母细胞是弥补这一差距的理想系统。我们和其他人已经证明，聚集体在卵子发生过程中积累。细胞质和细胞核中负责 RNA 转录和加工的核颗粒。三种RNA聚合酶自组装成聚集体，通常以RNA作为共聚集体。该提案的重点是核仁，一种重要的非膜结合颗粒，负责核糖体 RNA 的合成和加工、核糖体的制造、帮助介导细胞对压力和 DNA 损伤的反应，并适当地响应细胞对新物质的需求我们使用显微镜、免疫组织化学、生化分级、蛋白质。耗竭、蛋白质组和转录组分析，以了解积累、功能和稳定性我们利用了独特的优势来建立核仁的形式和功能。非洲爪蟾卵母细胞核的特性，包括大小、丰富的蛋白质和 RNA 含量以及易于分离这些发现将影响我们对细胞如何维持的理解。蛋白质的天然形式和聚集形式之间的生理正常平衡。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The ABCF gene family facilitates disaggregation during animal development.

ABCF 基因家族促进动物发育过程中的分解。

DOI：
发表时间：
2020
期刊：
影响因子：
3.3
作者：
Skuodas, Sydney;Clemons, Amy;Hayes, Michael;Goll, Ashley;Zora, Betul;Weeks, Daniel L;Phillips, Bryan T;Fassler, Jan S
通讯作者：
Fassler, Jan S

Component analysis of nucleolar protein compartments using Xenopus laevis oocytes.

使用非洲爪蟾卵母细胞对核仁蛋白区室进行成分分析。