Residue-by-residue details of FUS protein phase separation and aggregation

FUS 蛋白相分离和聚集的逐个残基详细信息

基本信息

批准号：
10708849
负责人：
Nicolas Lux Fawzi
金额：
$ 38.45万
依托单位：
BROWN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-23 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10708849
关键词：
Amino Acids Amyotrophic Lateral Sclerosis Aromatic Amino Acids Automobile Driving Binding Biological Biological Assay Biological Models Biological Process Biophysics C-terminal Cations Cell Communication Cell fusion Cell model Cell physiology Cells Cellular Stress Characteristics Charge Chimeric Proteins Chromosomal translocation Complement Complex Computer Simulation Cytoplasmic Granules DNA Binding Domain DNA Damage DNA Polymerase II DNA Repair Data Disease Explosion Family Foundations Frequencies Frontotemporal Dementia Functional disorder Future Gene Expression Genetic Transcription Goals Human In Vitro Knowledge Lead Link Liquid substance Malignant Neoplasms Mediating Membrane Missense Mutation Modeling Modification Molecular Molecular Probes Mutation NMR Spectroscopy Neurodegenerative Disorders Neurons Nuclear Localization Signal Oncogenic Organelles Pathologic Pattern Phase Physiological Physiology Play Post-Translational Protein Processing Process Proteins RNA RNA Folding RNA Polymerase II RNA Processing RNA Recognition Motif RNA Splicing RNA metabolism RNA-Binding Proteins Resolution Role Signal Induction Structure TAF15 gene Techniques Testing Transcription Coactivator Transcriptional Activation Untranslated RNA Visualization Work beta pleated sheet cell behavior effective therapy familial amyotrophic lateral sclerosis frontotemporal lobar dementia amyotrophic lateral sclerosis fused in sarcoma in vivo insight leukemia multimodality novel therapeutics paralogous gene pharmacologic prevent protein structure response sarcoma simulation structural biology transcription factor

项目摘要

Project Summary RNA-binding proteins are essential components of numerous large complexes that carry out fundamental processes including transcription, splicing, and DNA repair. Many RNA-binding proteins possess regions predicted to be disordered based on low-complexity sequence characteristics that are critical to normal RNA- processing functions, but also drive aberrant protein assembly in various neurodegenerative disease and cancers. The molecular interactions and functional roles of these disordered domains remain incompletely characterized, especially in the context of disease. Fused in Sarcoma (FUS) is one of twenty-nine human RNA-binding proteins that contains both an essential disordered low-complexity domain (LC) with unusually low charged residue composition and a high frequency of aromatic amino acids as well as several RGG motif regions. Despite disorder, these domains are thought to facilitate interactions in normal RNA metabolism by forming dynamic associations, thereby enabling tunable, reversible spatial clustering. Yet, excessive self- association between FUS disordered domains is believed to result in the formation of pathological neuronal inclusions in sub-types of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), irreversible neurodegenerative diseases that lack effective treatments. Moreover, fusion of the FUS disordered domains to several DNA-binding domains through chromosomal translocations results in uncontrolled gene expression leading to a family of aggressive cancers. Though FUS has emerged as the primary model system for understanding biological phase separation, the contacts holding together FUS assemblies and their structures in physiology and disease are currently unknown because they are invisible to traditional techniques in structural biology. However, we have demonstrated that we can visualize dynamic assemblies of FUS with residue-level resolution. This project will apply advanced nuclear magnetic resonance spectroscopy, molecular simulation, and cell models of FUS function to 1) visualize the molecular contacts that mediate LLPS and in- cell interactions, 2) probe the molecular and cellular impact of LC and RGG mutations of FUS found in familial ALS, and 3) determine the functionally relevant atomic details of FUS complexes with RNA and the C-terminal domain (CTD) of RNA polymerase II associated with RNA processing and transcription. These studies of FUS assembly will provide necessary structure/function information on future pharmacological targets for inhibiting pathological protein associations in types of ALS, FTD, leukemia, and sarcoma. Furthermore, because FUS is only one of many essential RNA-binding proteins containing aggregation-prone low complexity domains, the results of the project will serve as a foundation for understanding an entire class of proteins and for correcting their dysfunctions in disease.

项目摘要 RNA结合蛋白是许多大型复合物的重要组成部分包括转录，剪接和DNA修复的过程。许多RNA结合蛋白具有区域预计基于低复杂性序列特征而被无序，这对于正常RNA至关重要处理功能，但也驱动各种神经退行性疾病中的异常蛋白质组装和癌症。这些无序结构域的分子相互作用和功能作用不完全特征是，尤其是在疾病的背景下。融合在肉瘤（FUS）中是29人之一 RNA结合蛋白既包含一个本质无序的低复杂性结构域（LC）低电荷的残留物组成和高频的芳族氨基酸以及几个RGG基序地区。尽管有混乱，但这些领域仍被认为可以促进正常RNA代谢的相互作用形成动态关联，从而实现可调的，可逆的空间聚类。然而，自我过多据信，FUS无序结构域之间的关联会导致病理神经元的形成肌萎缩性外侧硬化症（ALS）和额颞痴呆（FTD）的子类型的夹杂物，不可逆缺乏有效治疗的神经退行性疾病。此外，融合了Fus无序域与通过染色体易位的几个DNA结合结构域导致基因表达不受控制导致一个激进的癌症家族。尽管FUS已成为主要模型系统了解生物阶段分离，将FUS组件及其结构组合在一起的触点目前在生理和疾病中尚不清楚，因为它们对传统技术是看不见的结构生物学。但是，我们已经证明我们可以可视化FUS的动态组件残留级分辨率。该项目将采用晚期核磁共振光谱，分子模拟，FUS功能的细胞模型1）可视化介导LLP和In-的分子接触细胞相互作用，2）探测家族性的LC和RGG突变的分子和细胞影响 ALS和3）确定与RNA和C末端的FUS复合物的功能相关的原子细节 RNA聚合酶II的结构域（CTD）与RNA处理和转录相关。这些关于FUS的研究组装将提供有关抑制未来药理目标的必要结构/功能信息 ALS，FTD，白血病和肉瘤类型中的病理蛋白质关联。此外，因为FUS是只有许多基本的RNA结合蛋白中的一个含有聚集的低复杂性域，该项目的结果将成为理解整个蛋白质并纠正的基础他们的疾病功能障碍。

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）

Structure and position-specific interactions of prion-like domains in transcription factor Efg1 phase separation.

转录因子 Efg1 相分离中朊病毒样结构域的结构和位置特异性相互作用。

DOI：
10.1101/2023.11.09.566450
发表时间：
2023
期刊：
bioRxiv : the preprint server for biology
影响因子：
0
作者：
Wang,Szu-Huan;Zheng,Tongyin;Fawzi,NicolasL
通讯作者：
Fawzi,NicolasL

Structure and interactions of prion-like domains in transcription factor Efg1 phase separation.

转录因子 Efg1 相分离中朊病毒样结构域的结构和相互作用。

DOI：
10.1016/j.bpj.2024.01.030
发表时间：
2024
期刊：
Biophysical journal
影响因子：
3.4
作者：
Wang,Szu-Huan;Zheng,Tongyin;Fawzi,NicolasL
通讯作者：
Fawzi,NicolasL

Liquid-Liquid Phase Separation of the Intrinsically Disordered Domain of the Fused in Sarcoma Protein Results in Substantial Slowing of Hydration Dynamics.

DOI：
10.1021/acs.jpclett.3c02790
发表时间：
2023-12-14
期刊：
JOURNAL OF PHYSICAL CHEMISTRY LETTERS
影响因子：
5.7
作者：
Krevert, Carola S.;Chavez, Daniel;Chatterjee, Sayantan;Stelzl, Lukas S.;Puetz, Sabine;Roeters, Steven J.;Rudzinski, Joseph F.;Fawzi, Nicolas L.;Girard, Martin;Parekh, Sapun H.;Hunger, Johannes
通讯作者：
Hunger, Johannes