Cytoplasmic organization by phase separation

通过相分离进行细胞质组织

• 批准号: 10437670
• 负责人: Amy Susanne Gladfelter
• 金额: $ 30.41万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2023-04-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10437670
• 关键词: Address; Amino Acid Sequence; Architecture; Base Pairing; Binding; Biochemical Reaction; Biochemistry; Biophysics; Cell Cycle; Cell Death; Cell Polarity; Cell Shape; Cell division; Cell membrane; Cell physiology; Cells; Cellular biology; Cyclins; Data; Diffuse; Disease; Elements; Genetic; Goals; Health; Human; Human Pathology; In Vitro; Lead; Link; Lipid Bilayers; Location; Malignant Neoplasms; Mediating; Membrane; Messenger RNA; Modeling; Molds; Molecular; Molecular Genetics; Neurodegenerative Disorders; Normal Cell; Nucleic Acids; Output; Pathologic; Pathology; Pattern; Phase; Physical condensation; Physiological; Porosity; Property; Proteins; Publishing; RNA; RNA Sequences; RNA-Binding Proteins; Reaction; Role; Site; Specific qualifier value; Specificity; Structure; Surface Tension; System; Tertiary Protein Structure; Time; To specify; Translations; Viscosity; Work; base; biophysical model; cell growth; experimental study; high resolution imaging; human disease; imaging genetics; molecular dynamics; nanoscale; nuclear division; physical property; physical state; polyglutamine; protein protein interaction; protein structure; reconstitution; scaffold; simulation

Project Summary/Abstract (PI Gladfelter, AS) Cells must compartmentalize biochemistry in time and space. A newly appreciated mechanism of organization is biomolecular condensation. In many cases, condensates form via weak, multivalent interactions among disordered proteins and nucleic acids. These interactions determine the material states of condensates such as viscosity, surface tension and porosity, which in turn impact the concentrations, reaction and transport rates in, out and within condensates of key constituents. There are major gaps in understanding how cells control where condensates form, which molecules coassemble, and how condensate material state contributes to function. We discovered a physiological function for condensates in controlling nuclear division and cell polarity in the filamentous fungus, Ashbya gossypii. These condensates can control translation and are formed by an RNA-binding protein called Whi3 binding to target RNAs important for nuclear division (cyclins) and cell polarity (formins). The power of this cell system is that we can link physical properties and locations of condensates to functional outputs of protein translation, cell shape and nuclear division. The goals of the proposed work are to determine how structured elements in proteins, RNAs and cell membranes control the material state, location and function of condensates in the cell. We will determine how nanometer scale features of protein and RNA sequences promote mesoscale physical states of condensates to spatially pattern protein translation. We use an interdisciplinary suite of advanced imaging, genetic, biophysical and modeling approaches to tackle these fundamental open problems that not yet understood for any phase-separating system. Specifically, we will: Aim 1: Determine roles of hidden structured domains of proteins. We hypothesize that transiently ordered states promote specific protein-protein interactions and condensate material properties. Aim 2. Establish the architecture and function of RNA-based scaffolds. We hypothesize that mRNA forms a higher-order network using base-pairing that determines condensate properties. Aim 3: Delineate how membrane platforms control condensate assemblies. We hypothesize that endomembranes provide sites of assembly to specify the location of condensates. The proposed work will define how protein structure, RNA scaffolds and cell membranes are harnessed to control the properties, functions and locations of condensates in cells. The importance of condesates is underscored by numerous findings that link aberrant formation of condensates to multiple human diseases, including cancer and neurodegenerative diseases. While it is clear condensates undoubtably impact biochemistry, we do not yet understand how condensates actually contribute to normal cell function which is critical to understand how their malfunction leads to human pathologies.

项目摘要/摘要（Pi Gladfelter，AS） 细胞必须在时间和空间中划分生物化学。组织的新机制 是生物分子冷凝。在许多情况下，冷凝水通过弱的多价相互作用形成 蛋白质和核酸无序。这些相互作用决定了冷凝水的材料状态 作为粘度，表面张力和孔隙率，这反过来影响浓度，反应速率 关键成分的冷凝水内，外部和内部。理解细胞如何控制有主要差距 凝结形成的地方，分子凝结的位置，以及凝结物材料状态如何贡献 功能。我们发现了控制核分裂和细胞的冷凝物的生理功能 丝状真菌的极性，Ashbya Gossypii。这些冷凝水可以控制翻译并形成 通过一种称为WHI3与靶RNA结合的RNA结合蛋白对核分裂（细胞周期蛋白）和细胞很重要的RNA 极性（formins）。该单元系统的功能是我们可以将物理属性和位置联系起来 凝结与蛋白质翻译，细胞形状和核分裂的功能输出。目标的目标 建议的工作是确定蛋白质，RNA和细胞膜中的结构化元素如何控制 材料状态，凝结物在细胞中的位置和功能。我们将确定纳米量表 蛋白质和RNA序列的特征促进了冷凝物的中尺度物理状态，以空间模式 蛋白质翻译。我们使用高级成像，遗传，生物物理和建模的跨学科套件 解决这些基本的开放问题的方法，这些问题尚未理解任何阶段分离 系统。具体来说，我们将：目标1：确定蛋白质隐藏结构域的作用。我们 假设瞬时有序状态促进特定的蛋白质 - 蛋白质相互作用和凝结 材料特性。目标2。建立基于RNA的支架的结构和功能。我们假设 该mRNA使用碱基配对形成了一个高阶网络，该网络决定了凝结物特性。目标3： 描述膜平台如何控制冷凝水组件。我们假设该内膜 提供组装位点以指定冷凝水的位置。拟议的工作将定义蛋白质 利用结构，RNA支架和细胞膜来控制特性，功能和位置 细胞中的冷凝物。偏见的重要性被许多发现异常联系的发现强调了 形成了多种人类疾病的冷凝物，包括癌症和神经退行性疾病。 虽然清楚地凝结无疑会影响生物化学，但​​我们尚不了解如何冷凝水 实际上有助于正常的细胞功能，这对于了解其故障如何导致人类至关重要 病理。