Learning principles from the pyrenoid, a phase-separated organelle

学习类核蛋白（一种相分离细胞器）的原理

基本信息

批准号：
10544349
负责人：
Martin Casimir Jonikas
金额：
$ 50.44万
依托单位：
PRINCETON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-01-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10544349
关键词：
Activase Algae Amyotrophic Lateral Sclerosis Architecture Back Binding Binding Sites Biochemical Process Biological Biological Models Biology Biophysical Process Carbon Dioxide Cell physiology Cells Cellular Structures Chlamydomonas Chlamydomonas reinhardtii Chloroplasts Collaborations Coupling Cryo-electron tomography Cues Data Disease Energy-Generating Resources Engineering Ensure Enzymes Equilibrium Exclusion Experimental Models Fluorescence Recovery After Photobleaching Free Energy Functional disorder Gene Expression Genetic Materials Health Holoenzymes Image In Vitro Individual Learning Liquid substance Location Magic Malignant Neoplasms Measurement Mediating Membrane Metabolism Microscopic Modeling Molecular Multienzyme Complexes Mutation Nature Organelles Phase Physical condensation Physiological Processes Positioning Attribute Process Property Proteins RNA Regulation Ribosomes Ribulose-Bisphosphate Carboxylase Role Signal Transduction Source Specificity Starch Structure System Thylakoids Variant Visualization biological adaptation to stress biophysical model carbon fixation experimental study flexibility human disease in vivo interdisciplinary approach liquid dynamics model organism molecular dynamics physical property recruit repaired response sample fixation self assembly stoichiometry theories

Activase Algae Amyotrophic Lateral Sclerosis Architecture Back Binding Binding Sites Biochemical Process Biological Biological Models Biology Biophysical Process Carbon Dioxide Cell physiology Cells Cellular Structures Chlamydomonas Chlamydomonas reinhardtii Chloroplasts Collaborations Coupling Cryo-electron tomography Cues Data Disease Energy-Generating Resources Engineering Ensure Enzymes Equilibrium Exclusion Experimental Models Fluorescence Recovery After Photobleaching Free Energy Functional disorder Gene Expression Genetic Materials Health Holoenzymes Image In Vitro Individual Learning Liquid substance Location Magic Malignant Neoplasms Measurement Mediating Membrane Metabolism Microscopic Modeling Molecular Multienzyme Complexes Mutation Nature Organelles Phase Physical condensation Physiological Processes Positioning Attribute Process Property Proteins RNA Regulation Ribosomes Ribulose-Bisphosphate Carboxylase Role Signal Transduction Source Specificity Starch Structure System Thylakoids Variant Visualization biological adaptation to stress biophysical model carbon fixation experimental study flexibility human disease in vivo interdisciplinary approach liquid dynamics model organism molecular dynamics physical property recruit repaired response sample fixation self assembly stoichiometry theories

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项目摘要

PROJECT ABSTRACT Phase separation is an emerging organizing principle for intracellular biology. Processes that are now understood to exploit phase separation include storage of genetic material, gene expression, ribosome synthesis, signaling, stress response, and metabolism. While each phase-separating system has unique features, there are universal themes relevant to all such systems, including regulation of the phase boundary, the dynamics of mixing within and between condensates, and the interactions of condensates with their surroundings. To uncover general principles regarding these common themes, we focus on a well-suited model organism and system: the genetically tractable alga Chlamydomonas reinhardtii and its pyrenoid, a non-membrane bound, phase-separated organelle responsible for efficient carbon fixation. The pyrenoid offers many practical advantages: 1. its phase separation is driven by two well- characterized components, the rigid enzyme complex Rubisco and the flexible linker protein EPYC1, via a known specific binding interface; 2. the pyrenoid’s in vivo liquidity is reproduced in vitro with no energy source; 3. in vivo assembly/disassembly is controllable by external cues; and 4. the pyrenoid is singular, large, and stable enough to systematically investigate its functional interactions with other cellular components. Based on these advantages, the pyrenoid has already proven to be a source of many discoveries including the ability of a flexible multivalent linker to condense a rigid component, inheritance of non-membrane bound organelles by fission, specific recruitment via a conserved binding motif, and a magic-number effect. The key universal questions we will address with this system are: What is the role of the valence, strength, and spacing of the interacting motifs in determining condensate properties? How does the stability of small oligomers control phase boundaries? What keeps condensates in a liquid state? How do cells control the number, size, and location of condensates, including their relation to other cellular structures? Our approach will closely integrate theory and experiment, as providing fundamental answers to these questions requires a multidisciplinary approach that places specific data within a broad theoretical framework. We anticipate that our focus on underlying biophysical mechanisms will facilitate generalizability of our results to a wide range of phase-separated intracellular systems.

项目摘要相位分离是细胞内生物学的新兴组织原理。现在正在了解利用相分离包括遗传物质的储存，基因表达，核糖体合成，信号传导，应力反应和代谢。虽然每个相分开系统都有唯一的功能，存在与所有此类系统有关的通用主题，包括对阶段的调节边界，冷凝水内和之间混合的动力学以及冷凝水的相互作用与周围环境。要揭示有关这些共同主题的一般原则，我们专注于合适的模型有机体和系统：一般可拖动的藻类衣原体Reinhardtii和它的类球形是一种非膜结合的，相位分离的细胞器，负责有效的碳固定。类拟南芥具有许多实用的优势：1。其相位分离是由两个良好的驱动的表征成分，刚性酶复合物rubisco和柔性接头蛋白EPYC1，已知的特定绑定界面； 2。类体内流动性在体外复制而没有能量来源; 3。体内组装/拆卸由外部提示控制；和4。类拟南芥是奇异的，较大，足够稳定，可以系统地研究其与其他细胞的功能相互作用成分。基于这些优势，类似力类动物已经被证明是许多人的来源发现包括灵活的多价链接器凝结刚性组件的能力，继承通过裂变，通过构成结合基序进行的特定募集的非膜结合细胞器的构成和A 魔术效应。我们将使用此系统解决的主要通用问题是：角色是什么相互作用基序的价，强度和间距在确定冷凝物特性时？如何小寡聚物的稳定性是否控制阶段边界？是什么使液体中的冷凝水保持状态？细胞如何控制冷凝物的数量，大小和位置，包括它们与其他的关系细胞结构？我们的方法将紧密整合理论和实验，以提供基本这些问题的答案需要一种多学科的方法，将特定数据列入广泛理论框架。我们预计我们专注于潜在的生物物理机制将有助于我们的结果对各种相位分离的细胞内系统的概括性。