Sequence determinants of membraneless organelle rheology

无膜细胞器流变学的序列决定因素

• 批准号: 10275998
• 负责人: Benjamin S Schuster
• 金额: $ 37.14万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-15 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10275998
• 关键词: Amino Acid Sequence; Binding; Biological Process; Biology; Biophysics; Biotechnology; Blood; Cell Nucleolus; Cell physiology; Cytoplasmic Granules; Development; Disease; Foundations; Gel; Gene Expression Regulation; Germ; Goals; Health; Link; Liquid substance; Measures; Microscopy; Molecular; Mutagenesis; Nanotechnology; Nerve Degeneration; Organelles; Pathology; Phase; Phase Transition; Physiological; Process; Property; Protein Engineering; Proteins; Research; Rheology; Role; Solid; Subcellular structure; Toxic effect; Veins; Vision; Work; biological adaptation to stress; biophysical techniques; bone; nanoparticle; stress granule; targeted treatment

PROJECT SUMMARY/ABSTRACT The long-term vision of my lab is to elucidate biophysical principles of mesoscale assembly in biology and to harness those discoveries for biomedical and biotechnological advances. In that vein, the lab’s current focus is on elucidating principles underlying intracellular phase separation. Phase separation is a fundamental process that cells use to organize their myriad biomolecules into functional compartments, giving rise to membraneless organelles such as stress granules, germ granules, and the nucleolus, with important roles in gene regulation, stress response, and many additional essential functions. Conversely, aberrant phase transitions are associated with neurodegenerative and other diseases. The overall goal of this MIRA proposal is to establish how biomolecular sequence and composition determine the material properties of membraneless organelles, and how material properties contribute to biological function and misfunction. We will dissect the sequence features and molecular interactions that determine the rheology (liquid-like, solid-like, or a combination) of biomolecular condensates formed by phase separation of intrinsically disordered proteins. We will accomplish this using protein engineering, mutagenesis, microscopy, a suite of biophysical techniques, and nanotechnology, including the development of a toolbox of nanoparticle probes for measuring membraneless organelle rheology. We will then investigate how membraneless organelle rheology underlies biological function, focusing on germ granules, and we will investigate how perturbations to membraneless organelle rheology perturb biological function. Finally, we will examine whether certain sequence features are particularly prone to aberrant liquid-solid phase transitions and toxicity, indicating possible origins of pathology and targets for therapies. Together, this work seeks to provide a strong foundation for understanding the links between molecular sequence, rheology, and function of membraneless organelles in health and disease.

项目摘要/摘要 我的实验室的长期愿景是阐明生物学中尺度组装的生物物理原理，并 利用这些发现促进生物医学和生物技术的进步。在这种情况下，该实验室目前的重点是 关于阐明细胞内相分离的基本原理。相分离是一个基本的过程。 细胞用来将它们的无数生物分子组织成功能隔间，从而产生无膜 应激颗粒、胚芽颗粒和核仁等在基因调控中起重要作用的细胞器， 应激反应，以及许多额外的基本功能。相反，反常的相变是相关的。 患有神经退行性疾病和其他疾病。Mira提案的总体目标是确定如何 生物分子序列和组成决定了无膜细胞器的材料性质，以及 材料特性如何导致生物功能和功能失调。我们将对序列特征进行剖析 以及决定生物分子的流变性(类液体、类固体或其组合)的分子相互作用 由本质上无序的蛋白质相分离而形成的凝聚体。我们将使用以下工具来完成此任务 蛋白质工程、诱变、显微镜、一套生物物理技术和纳米技术，包括 用于测量无膜细胞器流变学的纳米颗粒探针工具箱的研制。我们会 然后研究无膜细胞器流变学是如何支持生物功能的，重点是细菌颗粒， 我们将研究无膜细胞器流变学的扰动如何扰乱生物功能。 最后，我们将检查某些序列特征是否特别容易出现液-固相异常 过渡和毒性，表明可能的病理起源和治疗目标。共同努力，这项工作 旨在为理解分子序列、流变学和 无膜细胞器在健康和疾病中的作用。