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

• 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
• 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' 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

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。 不受控制。不久前，蛋白质聚集在很大程度上与发病机制等同。更多的 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 记忆并自组装成细胞核和细胞质中关键的非膜结合颗粒。如果 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疾病，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 淀粉样蛋白的特征。聚集体可以大大增加局部浓度 聚集蛋白质，并且是不同结构状态之间平衡的一部分。 因此，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。有一个差距 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 体内。我们建议开始弥合差距的理想系统是青蛙的卵母细胞 非洲爪蟾。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。这 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 翻译能力。我们使用显微镜、免疫组织化学、生化分级分离、蛋白质 耗尽、蛋白质组和转录组分析，以了解积累、功能和稳定性 建立核仁形式和功能的聚集体。我们利用独特的优势 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 操纵来进行我们的研究。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

The ABCF gene family facilitates disaggregation during animal development

• DOI: 10.1091/mbc.e19-08-0443
• 发表时间: 2020-04
• 期刊: Molecular Biology of the Cell
• 影响因子: 3.3
• 作者: Sydney Skuodas;A. Clemons;Michael H. Hayes;A. Goll;B. Zora;D. L. Weeks;B. Phillips;J. Fassler

Component analysis of nucleolar protein compartments using Xenopus laevis oocytes.

• DOI: 10.1111/dgd.12794
• 发表时间: 2022-08
• 期刊: Development, growth & differentiation